Research Report 2009 - Florey Institute

Research Report 2009

Table of ContentsDirector’s Report 1Behavioural Neuroscience— Neural Plasticity 2— Molecular Neurobiology 8— Addiction Neuroscience 11Brain Development and Regeneration 14Clinical Trials and Neuroscience Trials Australia 18Cognitive Neuroscience 24Epilepsy— Epilepsy Imaging 28— In Brain Markers of Epilepsy 31— Ion Channels and Human Disease 35Genomic Disorders Research Centre 44Imaging— Neuroimaging and Neuroinformatics 48— MRI Development 54Multiple Sclerosis 62Neurodegeneration— Molecular Neuropharmacology 70— Neural Development and Regeneration 75— Neurodegeneration 78— Steroid Neurobiology 83Neuropeptides 86Neuro Research Services— Formerly Integrative Neuroscience Facility 96Statistics and Decision Support 98Stroke— Basic Science 100— Epidemiology and Public Health 103— Stroke Imaging and Ultrasound 108— Neurorehabilitation and Recovery 112Systems Neurophysiology— Brain Function in Health and Disease 118

Our MissionImproving life throughbrain researchOur VisionTo be recognised as aleading international brainresearch instituteOur ValuesInnovation and excellence,commitment and passionintegrity and rigour,collaboration and team workLeft and right: Construction imagesfrom the Parkville and Austin sites

Director’s ReportSummarising 2009, the three keywords I would use to describe theFlorey Neuroscience Institutes’(FNI) progress are collaboration,construction and amalgamation.CollaborationFNI chartered a new course in itsscientific vision and structure through2009. Through the formation of a facultyand the development of a scientific visionwe have moved a long way to breakingdown the divide between campusesand we have built collaboration throughresearch themes.Through consultation our scientific groupsamalgamated into 12 research divisions,and where possible, they have beenco-led by clinical and basic scientists tofoster problem solving. The divisionshave brought a sense of structure to thescientific output across FNI.The 2009 research divisions are:• Epilepsy• Stroke• Multiple Sclerosis• Neurodegeneration• Statistics & Informatics• Behavioural neuroscience• Imaging• Clinical trials• Systems neurophysiology• Cognitive neuroscience• Brain development & Regeneration• NeuropeptidesThe scientific vision and the establishmentof the divisional structure providedus scope to focus on future researchdirections. The forward strategy in thescientific vision includes the developmentor recruitment of teams in translationalresearch, computational neuroscience,stem cells and the genetics ofneurological disease.ConstructionFNI is partnering with the University ofMelbourne, the Mental Health ResearchInstitute, Austin Health and MelbourneHealth in the construction of two multimilliondollar research facilities. Thepurpose built centres will accommodate700 researchers and support staff, therebyforming the largest neuroscience groupin the southern hemisphere.The financial component of theconstruction phase was secured in 2009when the Commonwealth governmentannounced that FNI and its partnerswere successful in a $39.8 million grantapplication to the Health and HospitalsFund. The generous support of theCommonwealth and State governmentsas well as very significant private donorsmeans the project is fully funded andproceeding rapidly.The Parkville and Austin sites began theconstruction phase in 2009. The buildingsare designed to enhance collaboration,with open plan office spaces, casualmeeting rooms and a flexibleweb-enabled auditorium space.State-of-the-art animal facilities,integrated I.T. systems and theintroduction of new equipment willmake the task of doing the sciencemore productive for all.AmalgamationFurther work has been done in 2009to bring the three merging institutestogether under a single FNI umbrella.Apart from the establishment of researchdivisions that cut through scientificdivides, the organisation forged aheadwith administrative changes thatcreated efficiencies across in the areasof finance, human resources, marketing,communications and fundraising.The streamlined operation of ouradministrative teams has resulted inbetter service delivery for our scientists.Professor Geoffrey A DonnanMBBS MD FRACP FRCP (Edin)1

Behavioural Neuroscience— Neural PlasticityResearch OverviewThe human cerebral cortex containsbillions of neurons that areinterconnected by trillions of synapsesto form functional networks underlyingour most complex brain functions. It isonly after birth, when environmentalstimuli induce patterned neural activityvia the sensory pathways, that diversecortical functions begin to emerge. Oneof our general aims is to gain insight intothe way in which the genetic programregulating maturation and function ofthe cortex is dynamically moulded byenvironmental stimuli. Our explorationof normal brain development andfunction has provided new informationon mechanisms of experiencedependent‘plasticity’ in the healthycerebral cortex that may underlie higherbrain functions such as learning andmemory. One particular aspect of ourresearch is focused on understandinggene-environment interactions indisorders involving disruption of normalcortical development, plasticity and/orfunction, such as those associated withHuntington’s disease (involving abnormalcortical/striatal function and plasticity)and schizophrenia (involving abnormalcortical maturation and plasticity). Using arange of techniques, including molecularbiology, protein chemistry, neuroanatomyand mouse behavioural analysis, we areattempting to understand how genes andenvironment combine to affect specificaspects of brain and behaviour.Further study of the beneficial effectsof various environmental manipulationsmight provide direction for therapeuticapproaches of clinical value for braindiseases. Ongoing research aimsto investigate gene-environmentinteractions and their impact on brainfunction and behaviour, not only toelucidate disease mechanisms, butalso to use our approach as a toolto identify molecular targets for futuredevelopment of novel therapeutics.We have discovered molecular pathwaysthat process information from theenvironment and induce experiencedependentchanges in the structure andfunction of neurons in the cerebral cortex.Major Research ProjectsGene-environment interactions andmolecular mechanisms mediatingcognitive, psychiatric and motorsymptoms in Huntington’s diseaseTerence Pang, Thibault Renoir,Michelle Zajac, Xin Du, Leah Leangand Anthony HannanHuntington’s disease (HD) is a devastatingillness in which a movement disorder(including chorea), cognitive deficits(dementia) and psychiatric symptoms(including depression) progress for 10-20years after onset, inevitably leading todeath. HD is caused by an expansionof a trinucleotide (CAG) repeat in asingle gene that is autosomal dominant,and thus will be inherited by 50%of the offspring of sufferers. Classicneuropathology is characterised bydegeneration of the cerebral cortexand striatum. Based on its autosomaldominant pattern of inheritance, HDhas long been viewed as the epitome ofgenetic determinism, however, our workhas demonstrated that environmentalfactors play a significant role inmodulating the disease process. Utilisinga transgenic mouse model of HD, wehave demonstrated that environmentalenrichment (a paradigm for enhancedmotor, cognitive and sensory stimulation)delays the onset and progression ofmotor symptoms and degenerationof the cerebral cortex. This work, incollaboration with colleagues at OxfordUniversity, provided the first evidencethat onset and progression of HD couldin fact be modulated by environmentalfactors. Following on from this work, ourlaboratory members have undertakena large battery of mouse behaviouraltests using HD mice, characterising bothlearning and memory deficits (modellingdementia in HD) and depressivelikebehaviours (modelling affectivesymptoms in HD). Furthermore, we haveextended the original findings to showthat in addition to the beneficial effectson motor symptoms, environmentalstimulation can also ameliorate deficitsin learning and memory, as well as thedepressive behaviours in HD mice.To better understand the complex natureof gene-environment interactions, wehave begun to compare the effects ofcomplex sensory and motor stimulationwith simple voluntary physical exercise(wheel running) in mice. Our work ongene expression and profiling studiesare identifying novel molecular targetswhose regulation are disrupted in HDbut may mediate the beneficial effectsof environmental enrichment. We nowhave evidence for molecular changesin synapses (synaptic plasticity) whichoccur in distinct regions of the cerebralcortex, as well as other brain areas,after exposing mice to different formsof environmental stimulation. This linkbetween environmental stimulation andchanges in the synaptic connectionsbetween specific neuronal populations,gives us new insight into the way inwhich environmental factors may delayonset and progression of HD and relatedneurodegenerative diseases.One implication of these findings isthat all brain disorders, even hereditaryones like HD, involve gene-environmentinteractions, and this idea has inspirednew international research on the roleof environmental factors in other braindiseases, including Alzheimer’s disease.Our research has provided new insightsinto how genes and environment affectbrain function and dysfunction, andhence normal and abnormal behaviour.Our development of novel experimentalmodels of gene-environment interactionswill have far-reaching implications forthe understanding and treatment ofbrain disorders. The establishment andintegration of genetic and environmentalmodels will facilitate the screening ofnovel therapeutic interventions targetinga wide range of genetically mediated,and environmentally modulated,brain diseases.Molecular mediators and environmentalmodulators in a transgenic mousemodel of depressionThibault Renoir, Terence Pang,Xin Du, Michelle Zajac, Christina Mo,Grace Chan, Andrew Argyropoulosand Anthony HannanDepression is the most commonpsychiatric disorder found in Huntington’spatients, and occurs at early stages inthese patients and at a much higherrate than the general community. Wehave recently discovered that we canaccurately model depression-likebehaviours in HD mice, and that theseprecede cognitive deficits and motor2

abnormalities. We are now in theprocess of investigating the use of otherantidepressants to treat the depressivelikebehaviours in HD mice, as well asinvestigating the impact of those drugson the brain at a molecular level, in theattempt to understand how changes ingene expression and molecular signalling(for example those involving theserotonergic system and neurotrophins)is translated to the behavioural level.We have found that environmentalenrichment, which induces increasedmental stimulation and enhancedphysical exercise, has antidepressant-likeeffects in the HD mice. This is associatedwith specific changes in serotonin (5-HT)receptors at both mRNA and proteinlevels, as well as cellular, physiologicaland behavioural effects. These findingsnot only have implications for HD butalso for our understanding of, andfuture development of better treatmentsfor, depression.The function of adult hippocampalneurogenesis in the healthy anddiseased brainMark Ransome and Anthony HannanIn addition to altered synaptic plasticity,we also have evidence for alteredcellular plasticity and have demonstratedthat HD mice have deficits in adultneurogenesis (the birth of new neurons)within the hippocampus. A deficit inneurogenesis has been postulatedas an underlying cause of depression.Following our discovery, with colleaguesat Cambridge University, that transgenicHD mice have decreased hippocampalcell proliferation, we hypothesised thatdecreased hippocampal neurogenesismay contribute to depressive symptomsand cognitive decline in HD. Selectiveserotonin reuptake inhibitor (SSRIs)prescribed for the treatment of clinicaldepression, are known to increasehippocampal neurogenesis in wildtypeanimals. We have discovered thathippocampal-dependent cognitive anddepressive-like behavioural symptomsoccur in HD mice. We are now examininghow the HD gene mutation changesspecific molecules in the dentate gyrusof the hippocampus, and how this mightimpact on adult neurogenesis. We arealso examining how environmentalstimuli, including increased cognitivestimulation and physical exercise,modulate adult neurogenesis in thehealthy and diseased hippocampus.Mechanisms of postnatal maturationand cellular plasticity in the cerebralcortex and their dysfunction in thepathogenesis of schizophreniaEmma Burrows, Elizabeth Manning,Mark Ransome and Anthony HannanDuring important postnatal periods, thebrain is particularly open to influencefrom environmental factors, withexternal inputs modifying structural andmolecular development. Examination ofthe molecular mechanisms underlyingthe interactions between environmentalfactors and the genetic determinants ofdevelopment allows us to understandthe complex pathways involved inboth normal and abnormal maturationprocesses. We have identified specificmolecular pathways that lie at theintersections of environmental andgenetic inputs. These pathways involveglutamate receptors, G proteins,regulators of G protein signalling, anddownstream second messengers, whichprocess information derived from thestimulation of the sensory organs andinduce activity-dependent changes inthe structure and function of neuronsin sensory cortex. We have used in situhybridisation and Western analysis tocharacterise spatiotemporal expressionpatterns of the genes during criticalperiods of developmental plasticity.Correlations between expressionpatterns and these critical periods insomatosensory and visual periods, as wellas other brain areas such as hippocampus,have been followed up by comparisonof normal mice with knockout littermates,in which one of the genes of interest hasbeen inactivated.Schizophrenia is a debilitatingdisorder that is thought to result fromcomplex interactions between multiplesusceptibility genes and environmentalinfluences, during postnatal and laterdevelopmental periods. Our studiesof the pathways that mediate geneenvironmentinteractions in developmentare intrinsically linked to the theories ofschizophrenic aetiology, and provide apowerful model for investigating thisdisorder. We are investigating whetherthe abnormal cortical developmentseen in schizophrenia is related todisruption of these molecular signallingpathways, and whether positive ornegative environmental influences canmodulate the resulting symptomatology.Mice deficient in phospholipase C-ß1(PLC-ß1) and the metabotrobic glutamatereceptor 5 (mGluR5), key genes inpathways of experience-dependentsynapse plasticity, display behaviouralabnormalities that show striking parallelsto the symptoms of schizophrenia. Wehave built a comprehensive profile ofthese behavioural symptoms, conductinga battery of tests to elucidate thefunctional domains affected by the genedeletions. We also seek to elucidate themechanisms by which environmentalfactors and pharmacotherapy canmodulate plasticity processes, and howthese modulate the behavioural andmolecular changes observed in theknockout mice. We have shown thatcommonly used pharmacotherapiesfor schizophrenia can reverse severalof the behavioural abnormalities inmGluR5 knockout mice, strengtheningthe validity of this approach to modellingschizophrenic symptoms. Recentimportant research from our laboratorydemonstrated that environmentalenrichment can reverse the schizophrenialikesymptoms in PLC-ß1 deficient mice.This finding highlights the importanceof behavioural interventions in diseasetreatment, and reinforces the hypothesisthat this signalling pathway may actas a point of intersection for geneticand environmental risk factors forschizophrenia. Identification of themolecular targets of environmentalfactors may present novel targets fordrug development.Other Research ProjectsThe influence of environmental factorson a mouse model of Rett syndromeand autismMari Kondo, Monique Howard andAnthony Hannan, in collaboration withGreg Pelka and Patrick Tam (Children’sMedical Research Institute, Sydney)Rett syndrome is an X-linked disorderinvolving mutations in the methyl-CpGbindingprotein Mecp2. It is characterisedby regression of motor and cognitivelearning early in life, leading to profoundautism-like behaviour, cognitiveimpairment and motor dysfunction.Abnormalities in neuronal structure andconnectivity have been observed in thedisorder, in addition to changes in theexpression of key regulatory proteins3

Behavioural Neuroscience–— Neural Plasticityincluding BDNF. The symptomatology ofindividuals with mutations in the Mecp2gene varies considerably, indicating thatpotential environmental factors maymodulate disease phenotype. Mutationsin Mecp2 have also been found to onegenetic factor in cases of autism, anothermajor neurodevelopmental disorderthat is thought to be caused by multiplegenetic and environmental factors. Weare investigating the interactions betweenthis genetic mutation and environmentalstimuli using a mouse knockout modelof the disorder. Behavioural, cellular andmolecular correlates of these changesare being assessed, and we are testingthe hypothesis that environmentalenrichment will ameliorate the severityof symptoms.Gtf2ird1 function in vivo – insightsinto Williams Beuren SyndromeMonique Howard and Anthony Hannan,in collaboration with Edna Hardeman(Children’s Medical Research Institute andUniversity of New South Wales, Sydney)Williams Beuren Syndrome (WBS) is amultifactorial neurodegenerative diseasepresent in up to 1 in 7,500 live births.Patients have distinct cognitive andbehavioural phenotypes including mild tomoderate intellectual learning disabilities,visuospatial learning difficulties, anxiety,attention deficit hyperactivity disorderand overfriendliness. WBS is caused bythe hemizygous deletion of a region ofchromosome 7 containing up to 28 genes.One gene in this region that is thoughtto contribute to some of the cognitiveand behavioural deficits is the GeneralTranscription Factor 2 I-repeat domain1 (Gtf2ird1). We are using a mousewith a deletion in the Gtf2ird1 gene toinvestigate the function of this genein vivo, providing us with informationabout both the manifestation of WBS, inaddition to aiding our understanding ofthe role of genes in neurodevelopmentaffecting cognitive and behavioural traits.Staff ListFellows, Senior Research Officers andResearch OfficersAnthony Hannan BSc (Hons) PhD (Syd)Terence Pang BSc (Hons) PhD (Melb)Mark Ransome BSc, MSc, PhD (Melb)Thibault Renoir BSc (Hons) PhD (Paris)Laura GrayBSc (Hons) PhD (Melb) (until March 2008)Caitlin McOmishBSc (Hons) PhD (Melb) (until April 2008)Jess NithianantharajahBSc (Hons) PhD (Melb) (until May 2008)Research AssistantsMonique Howard BSc (Hons) (La Trobe)Leah Leang BSc (Hons) MSc (ANU)StudentsTerence Pang BSc (Hons) PhD (Melb)(PhD thesis submitted September 2008)Emma Burrows BSc (Hons) (Melb)Michelle Zajac BSc (Hons) (Melb)Mari Kondo BSc (Hons) (Macquarie)Xin Du BSc (Hons student 2008;PhD student from 2009)Christina MoBSc (Hons)(UWA) (PhD student from 2010)Yanting Choong BSc (Hons student 2008)Elizabeth Manning BSc (Hons student 2009;PhD student from 2010 at MHRI/FNI)Andrew Argyropoulos BSc (Hons student2010)Grace Chan AMS studentYoshiko Shikano AMS studentConferences andPresentations (2008-2009)NationalAnthony HannanHuntington’s Research Group of VictoriaAnnual Scientific Meeting, Melbourne,2008Invited seminars:Australian Society for NeuroscienceAnnual Meeting, NeurogenesisSymposium, Canberra (2009)International Society for AffectiveDisorders meeting, BDNF Symposium,Brisbane (2009)School of Medicine, University of Sydney,Sydney (2009)National Centre for Adult Stem CellResearch, Eskitis Institute, GriffithUniversity, Brisbane (2009)School of Psychiatry, University ofNew South Wales, Sydney (2009)Genomic Disorders Research Centre,University of Melbourne (2009)Department of Pharmacology,University of Melbourne (2009)Cognitive Science Forum,University of Melbourne (2009)Department of Microbiology andImmunology, University of Melbourne(2009)School of Medical Sciences,University of Sydney (2009)Plenary Lecture, Academic Unit forPsychiatry of Old Age, University ofMelbourne, 14th Annual Symposium,Melbourne (2009)Department of Anatomy and Cell Biology,University of Melbourne (2009)Department of Pathology,University of Melbourne (2009)Florey Neuroscience Institutes AnnualNeuroscience Symposium, Melbourne(2009)Invited Lecturer, Neurogenetics on theApple Isle, Australian Society for MedicalResearch (ASMR) National ScientificCongress, Hobart (2009)4

Symposium Speaker, AustralasianSociety for Psychiatric Research Meeting,Canberra (2009)National Stroke Research Institute / BrainResearch Institute seminar, Melbourne(2008)Department of Biochemistry, Bio 21Institute, University of Melbourne (2008)Monash Neuroscience Seminar, MonashUniversity, Melbourne (2008)Plenary Lecture, Australian and NewZealand Society for Geriatric MedicineMeeting, Melbourne (2008)National Conference of the AustralianHuntington’s Disease Association,Adelaide (2008)Australasian Society for PsychiatricResearch Meeting, Newcastle (2008)3rd Frontier Technologies in NervousSystem Repair Workshop, Mt Lofty, SouthAustralia (2008)Australian Society for NeuroscienceAnnual Meeting, Plasticity Symposium,Hobart (2008)Terence PangHuntington’s Research Group ofVictoria Annual Scientific Meeting,Melbourne, 2008Michelle ZajacHuntington’s Research Group ofVictoria Annual Scientific Meeting,Melbourne, 2008Emma BurrowsNeurogenetics on the Apple Isle,Australian Society for Medical Research(ASMR) National Scientific Congress,Hobart (2009)Australian Society for NeuroscienceAnnual Meeting Hobart (2008)Mari KondoEpigenetics Conference,Melbourne, 2009Australian Society for NeuroscienceAnnual Meeting Hobart (2008)Xin DuHuntington’s Research Group ofVictoria Annual Scientific Meeting,Melbourne, 2008InternationalAnthony HannanFederation of European NeuroscienceSocieties, Geneva, Switzerland, 2008Society for Neuroscience, Washington,DC, USA, 2008Invited seminars:Regenerative Medicine, Pfizer,Cambridge, UK (2009)Department of Experimental Psychology,University of Cambridge, UK (2009)Department of Experimental Psychology,University of Oxford, UK (2009)European COST Meeting onneurodegeneration and regeneration,Antwerp, Belgium (2009)Institute of Neuroscience and Physiologyat Sahlgrenska Academy, Göteborg,Sweden (2009)Workshop Lecture, 4th European Societyfor Neurochemistry (ESN) Conferenceon Advances in Molecular Mechanismsof Neurological Disorders, Leipzig,Germany (2009)Plenary Lecture, Australasian WinterConference on Brain Research,Queenstown, New Zealand (2009)Videoconference seminar to nationalmeeting of Huntington Society of Canada(2008)4th International NeuroacanthocytosisSymposium, London/Oxford, UK (2008)Terence PangSociety for Neuroscience, Washington,DC, USA, 2008Hereditary Disease Foundation HD2008Meeting, Cambridge, MA, USA (2008)Emma BurrowsFederation of European NeuroscienceSocieties, Geneva, Switzerland, 2008Michelle ZajacWorld Congress of Huntington’s Disease,Vancouver, Canada, 2009Society for Neuroscience, Washington,DC, USA, 2008Mari KondoInternational Rett Syndrome Meeting,Chicago, USA, 2009Federation of European NeuroscienceSocieties, Geneva, Switzerland, 2008RIKEN Summer Brain Science School,Japan, 2008AwardsTerence PangPrize for Best Student Talk at theHuntington’s Research Group ofVictoria Annual Scientific Meeting,Melbourne, 2008Mark RansomeNHMRC Biomedical Fellowship,awarded in 2009Michelle ZajacWorld Federation of NeurologyTravel Award for the World Congressof Huntington’s Disease, Vancouver,Canada, 2009Emma BurrowsFNI Award for 3rd Year Student Talk, 2008Mari KondoFNI Student AwardXin DuMelbourne Research ScholarshipChristina MoAustralia Postgraduate AwardMajor Collaborative LinksNationalA/Prof Gary EganHoward Florey InstituteMagnetic Resonance Imaging of amouse model of Huntington’s diseaseProf Patrick Tam and Dr Greg PelkaChildren’s Medical Research Instituteand University of Sydney, SydneyCharacterisation of a mouse modelof Rett syndrome5

Behavioural Neuroscience— Neural PlasticityProf Edna Hardeman andDr Stephen PalmerUniversity of New South Wales, SydneyCharacterisation of a mouse model ofWilliams syndromeA/Prof Maarten van den BuuseMental Health Research Instituteof VictoriaPre-pulse inhibition in a mouse modelof schizophreniaA/Prof Brian Dean andDr Elizabeth ScarrMental Health Research Instituteof VictoriaNeuronal signalling proteins inschizophreniaDr Erica FletcherDepartment of Anatomy andCell Biology, University of MelbourneRetinal abnormalities in a mousemodel of HDA/Prof Xiao-Jun DuBaker-IDI, MelbourneCardiovascular abnormalities in a mousemodel of HDInternationalProf Richard Faull andDr Michelle GlassUniversity of Auckland, New ZealandCannabinoid receptors in Huntington’sdiseaseA/Prof Jang-Ho ChaMassachusetts General Hospital,Harvard University, Boston, USAActivity-dependent neurotransmitterreceptor expression in Huntington’sdiseaseDr Ghazaleh Sadri-VakiliMassachusetts General Hospital,Harvard University, Boston, USAEpigenetics in the healthy anddiseased brainA/Prof Ruth Luthi-CarterBrain Mind Institute, Swiss FederalInstitute of Technology, Lausanne,SwitzerlandsMolecular correlates of environmentalenrichment in Huntington’s diseaseDr Caitlin McOmishColumbia University, USABehavioural and pharmacologicalanalysis of a model of schizophreniaPublications (2008-09)Research ArticlesBenn, C.L., Luthi-Carter, R., Kuhn, A.,Sadri-Vakili, G., Blankson, K.L., Dalai, S.C.,Goldstein D.R., Spires, T.L., Pritchard, J.,Olson, J.M., van Dellen, A., Hannan, A.J.and Cha, J.-H.C. (2010) Environmentalenrichment reduces neuronalintranuclear inclusion load but has noeffect on mRNA expression in a mousemodel of Huntington’s disease. Journalof Neuropathology and ExperimentalNeurology [7 Jul 2010, Epub aheadof print].Dowie, M.J., Howard, M.L., Nicholson,L.F., Faull, R.L., Hannan, A.J. and Glass,M. (2010) Behavioural and molecularconsequences of chronic cannabinoidtreatment in Huntington’s diseasetransgenic mice. Neuroscience[1 Jul 2010, Epub ahead of print].Zajac, M.S., Pang, T.Y.C., Wong, N.,Weinrich, B., Leang, L.S.K. , Craig, J.M.,Saffery, R.and Hannan, A.J. (2009) Wheelrunning and environmental enrichmentdifferentially modify exon-specific BDNFexpression in the hippocampus of wildtypeand pre-motor symptomatic maleand female Huntington’s disease mice.Hippocampus 20:621-636.Dowie, M.J., Bradshaw, H.B., Howard,M.L., Nicholson, L.F.B., Faull, R.L.M.,Hannan, A.J. and Glass, M. (2009)Cannabinoid system abnormalitiesprior to motor onset in a transgenicmouse model of Huntington’s disease.Neuroscience 163:456-65.Gray, L., McOmish, C.E., Scarr, E., Dean,B. and Hannan, A.J. (2009) Sensitivityto MK-801 in phospholipase C-ß1knockout mice reveals a specific NMDAreceptor deficit. International Journal ofNeuropsychopharmacology [23 Feb 2009,Epub ahead of print].Nithianantharajah, J., Vijiaratnam, N.,Barkus, C., Clement, O. and Hannan, A.J.(2009) Modeling brain reserve:Experience-dependent neuronal plasticityin healthy and Huntington’s diseasetransgenic mice. American Journal ofGeriatric Psychiatry 17: 196-209 (invitedresearch article for special issue).6

Pang, T.Y.C., Du, X., Zajac, M.S.,Howard, M.L. and Hannan, A.J. (2009)Altered serotonin receptor expressionis associated with depression-relatedbehavior in the R6/1 transgenic mousemodel of Huntington’s disease. HumanMolecular Genetics 18: 753-766.McOmish, C.E., Burrows, E., Howard, M.and Hannan, A.J. (2008) PLC-ß1 knockoutmice as a model of disrupted corticaldevelopment and plasticity: behavioralendophenotypes and dysregulation ofRGS4 gene expression. Hippocampus18: 824-834.Gray, L., Howard, M., Scarr, E., van denBuuse, M., Dean, B. and Hannan, A.J.(2008) Clozapine reverses schizophreniarelatedbehaviours in the metabotropicglutamate receptor 5 knockout mouse:association with N-methyl-D-asparticacid receptor upregulation. InternationalJournal of Neuropsychopharmacology12: 45-60.Kondo, M., Gray, L., Pelka, G.J.,Christodoulou, J., Tam, P.P.L. andHannan, A.J. (2008) Motor deficitsin a Rett syndrome mouse model areameliorated by environmental enrichment.European Journal of Neuroscience 27:3342-3350.Stam, N., Nithianantharajah, J., Howard,M.L., Atkin, J.D., Cheema, S.S. andHannan, A.J. (2008) Sex-specificbehavioural effects of environmentalenrichment in a transgenic mouse modelof amyotrophic lateral sclerosis. EuropeanJournal of Neuroscience 28: 717-723.Gray, L., McOmish, C.E., Howard, M.and Hannan, A.J. (2008) Behaviouraleffects of the clozapine derivative,N-desmethylclozapine, in a geneticmouse model of schizophrenia.Behavioural Pharmacology 19: 543-547(invited research article for special issue).Gibbons, A.S., Scarr, E., McOmish, C.,Hannan, A.J., Thomas, E. and Dean, B.(2008) RGS4 expression is not altered inthe prefrontal cortex of schizophrenics.Australian and New Zealand Journal ofPsychiatry 42: 740-745.McOmish, C.E., Burrows, E., Howard,M., Scarr, E., Kim, D., Shin, H.-S., Dean,B., van den Buuse, M. and Hannan, A.J.(2008) Phospholipase C-ß1 knockoutmice exhibit endophenotypes modellingschizophrenia which are rescued byenvironmental enrichment and clozapineadministration. Molecular Psychiatry13: 661-72.Nithianantharajah, A.J., Barkus, C.,Murphy, M. and Hannan, A.J. (2008)Gene-environment interactionsmodulating cognitive function andmolecular correlates of synaptic plasticityin Huntington’s disease transgenic mice.Neurobiology of Disease 29:490-504.van Dellen, A., Cordery, P.M., Spires, T.L.,Blakemore, C. and Hannan, A.J. (2008)Wheel running from a juvenile age delaysonset of specific motor deficits but doesnot alter protein aggregate density in amouse model of Huntington’s disease.BMC Neuroscience 9: 34.Review Articles and Book ChaptersHannan, A.J. (2009) Tandem repeatpolymorphisms: modulators of diseasesusceptibility and candidates for ‘missingheritability’. Trends in Genetics 26: 59-65[24 Dec 2009, Epub ahead of print].Hannan, A.J. Gene-environmentinteractions in neurodegenerativediseases. In: Encyclopedia ofEnvironmental Health. Elsevier, Oxford(invited book chapter, in press).Hannan, A.J. (2010) Environmentalenrichment and gene-environmentinteraction in mouse models of braindisorders (Chapter 11: pp201-216) In:Transgenic and Mutant Tools to ModelBrain Disorders, Editors: A.V. Kalueff, C.L.Bergner. Humana Press, New Jersey(invited book chapter).Nithianantharajah, J. and Hannan, A.J.(2009). The neurobiology of brain andcognitive reserve: Mental and physicalactivity as modulators of brain disorders.Progress in Neurobiology 89:369-82.Pang, T.Y.C., Chiu, E., Chua, P., Georgiou-Karistianis, N., and Hannan, A.J. (2009)Depression in Huntington’s disease.Frontiers in Neuroscience 3: 238-239.Hannan, A.J. (2009) Towards a therapyfor Huntington’s disease (Commentaryon Giampà et al.). European Journal ofNeuroscience 29: 901.Fondon, J.W., Hammock, E.A.D., Hannan,A.J. and King, D.J. (2008) Simplesequence repeats: Genetic modulatorsof brain function and behavior. Trends inNeurosciences 31: 328-334.Laviola, G., Hannan, A.J. Macrì, S.,Solinas, M. and Jaber, M. (2008) Effects ofenriched environment on animal modelsof neurodegenerative and psychiatricdisorders. Neurobiology of Disease 31:159-168.Bohanna, I., Georgiou-Karistianis, N.,Hannan, A.J. and Egan, G.F. (2008)Magnetic Resonance Imaging asan approach towards identifyingneuropathological biomarkersfor Huntington’s disease.Brain Research Reviews 58: 209-225.Hannan, A.J. (2008) EuropeanNeuroscience – Sixth Biennial FENSForum. IDrugs 11: 643-645.7

Behavioural Neuroscience— Molecular NeurobiologyResearch OverviewThe Molecular Neurobiology laboratoryhas generated a number of geneknockout and knockin animal modelsdesigned to investigate brain diseases.These include D1 dopamine receptorknockout; four D1 receptor expressingcell ablation mutants in which (1) the cellsare killed globally throughout the brain or(2) from the cortex alone starting in earlydevelopment and (3) from the cortex andbasal ganglia or (4) basal ganglia alonebeginning in the post-natal brain andcontinuing throughout adulthood; and4 neuronal nicotinic receptor knockoutand Autosomal Dominant NocturnalFrontal Lobe Epilepsy (ADNFLE) modelS248F knockin lines. The behavioural,neuropharmacological, neuroanatomicaland neurophysiological characterisationof these models is contributing toour fundamental understandingof a number of diseases includingHuntington’s disease, nigrostriataldegeneration, dystonia and ADNFLEand drug addiction. Mechanisms ofneurodegeneration including oxidativestress, apoptosis, glial mediated neuronaldamage, stem cell proliferation, andcellular remodelling responses of thediseased brain to precise molecularlesioning are being studied in an effortto design effective therapeutic strategies.The nicotinic receptor knockout andknockin mutants generated by thegroup provide insights into models ofaltered nicotinic function in the brain thatmay ultimately explain mechanisms ofdystonia, nicotine addiction and nicotine/dopamine interactions. The laboratoryblends expertise in molecular and stemcell biology with clinical paradigmsto design research projects that willultimately impact on our understandingof mechanisms of neurodegenerativedisease. Clinical projects have focused ondeveloping strategies for early diagnosisof a range of neurodegenerativediseases presenting to clinicians as earlyimpairments of cognitive functions andParkinson’s disease.Major Research ProjectsBehavioural and anatomicalcharacterisation of mutant mice withprogressive ablation of D1 dopaminereceptor-expressing cellsHelena Kim, Luning Jiang, Ilse Gantois,Andrew J. Lawrence, Ann Graybiel,Satoshi Goto, John L. Waddingtonand John DragoWe have shown that ablation of D1+ cellsin the striatum gives rise to Parkinsonianmotor symptoms with impairments in gaitand balance and abnormal oral functionsbut no limb dystonia whereas cortical D1+cell ablation gives rise to limb dystoniawithout gait or oral function abnormality.Huntington disease involves the lossof both dopamine responsive striatalmedium spiny neurons as wellas cortical neurons. Clinical featuresinclude hyperkinetic involuntarymovements, gait and orofacialimpairments, cognitive deficits, andpsychosis, mood and anxiety disorders.We utilized the Cre-LoxP system togenerate a transgenic mouse line withselective ablation of D1 dopaminereceptor (Drd1a)-expressing striatalneurons (using a DARPP-32/Cre line).In this line, atrophy and astrogliosis ispresent in the striatum but not in thecortex. Drd1a, substance P, and dynorphinmRNA expression is reduced uniformlythroughout the entire rostrocaudal extentof the dorsal striatum, while striatal D2dopamine receptor and enkephalinmRNA is up-regulated. Mutant mice havereduced bodyweight and display motordeficits consistent with a parkinsonianphenotype including bradykinesia,impaired motor coordination, and a shortstepping gait. Ethological assessmentidentifies impairments in orofacialmovements of sifting and chewing.Mutant mice display reduced anxiety-likebehaviours but normal spatial workingmemory and surprisingly do not displaytail suspension limb dystonia. We usedD1-EGFP mice backcrosses to verifythat loss of D1+ cells was confinedto the striatum. NPY and cholinergicinterneurons were increased in thestriatum but cortical interneurons wereunchanged. The model demonstratesthat loss of Drd1a-expressing striatalneurons alone is sufficient to produceorofacial abnormalities and a robustParkinsonian motor phenotype.In addition, transgenic mice harboringsilenced diphtheria toxin gene underthe Drd1a gene were bred with Emx-1 Cremice to generate Emx-1 MUT animalsthat lacked Drd1a-expressing corticalpyramidal cells. Changes to striatal Drd1aexpression and other striatal markerswere not detected in the mutant mice.Adult Emx-1/MUT mice were hyperactive,displayed prominent forelimb clasping,poor rotarod performance and hadnarrow forelimb posturing. In addition,mutant animals also displayed increasedanxiety as well as impaired memory.Both motor- and somatosensory- cortexof Emx-1 MUT mice were reduced inthickness, particularly in the deepercortical layers compared with the controls.This correlated strongly with the loss ofDARPP-32-expressing cells in the sameregion. Older Emx-1 MUT mice displayedhindlimb dystonia in addition to forelimbdystonia. This study underscores theprimary role of cortical Drd1a expressingcells in motor control and cognition.Evaluation of the regenerative capacityand origin of adult dopaminergicneurons: an in-vitro model of ratmesencephalon derived adultdopaminergic neuronsShohreh Majd, Clare Parish,Arthur Smardencas and John DragoAn in-vitro rodent culture system for adultdopaminergic neurons was establishedto study transynaptic effects of striatalneuronal loss on dopaminergic neuronsas our in-vivo experiments had showna decrease in the level of striatal &cortical dopamine levels in DARPP-32/Cre MUT mice without primary lossof dopaminergic cells and a decreasein striatal DA levels in CamK2a/CreMUT mice. This suggests that impairedstriatonigral and ventrostriatal-VTA inputsimpact on DA cell activity. This can onlybe studies in individual cells. A culturesystem was therefore established inwhich adult brain derived DA neurons areisolated and grown in long term culture.In this study, we sought to establisha culture aiming to promote thesurvival and regeneration of adult ratmesencephalon derived dopaminergicneurons. We investigated the role ofFGF2, BDNF, TGFß3, GDNF and dbcAMP.The proportion of total neurons with adopaminergic phenotype after 8 days invitro was 1.5% in cultures maintained in8

FGF2, BDNF and GDNF. Dopaminergicneurons had long arborized processes.dbcAMP promoted neurite extensionbut did not enhance cell survival.Dopaminergic neurons derived fromBrDU incorporating dividing cells wereidentified in early cultures but werenot seen in long term cultures. Thisstudy shows that long term culturesof dopaminergic neurons can beestablished from the mature rat brainand dopamineregic neurons can bemanipulated by epigenetic factors suchas the growth factor and intracellularcAMP levels.Testing an algorithm to diagnoseAlzheimer’s and non-Alzheimer’sdementia at presentation with mildcognitive impairmentJohn Drago in collaboration withColin Masters, Christopher Rowe,Victor Villemagne and Qiao-Xin LiA working clinical diagnosis of mildcognitive impairment (MCI) can be madeat presentation with memory impairment.According to the Peterson criteria, MCIis a change in memory corroborated byan informant accompanied by objectiveevidence of impairment with preservationof activities of daily living. In makinga diagnosis of MCI, secondary (andpotentially reversible) causes of dementiaare excluded by clinical assessment anda standard dementia screen. Secondarycauses that must be excluded includestructural brain lesions such as subduralhaematoma, cerebral tumour andnormal pressure hydrocephalus, CNSinfections, metabolic abnormalities,severe depression and alcohol abuse.Amnestic-MCI as defined by thesecriteria is a significant medical entitybecause it may convert to a numberof fatal neurodegenerative diseases ofwhich dementia is a major feature. Therate of conversion of amnestic-MCI toAlzheimer’s disease (AD) (the largestsingle group) or other non-AD formsof dementia such as vascular dementia(VsD), Lewy body dementia (DLB),fronto-temporal dementia (FTD) orParkinsonian syndromes [corticobasaldegeneration (CBD), progressivesupranuclear palsy (PSP)], varies betweenstudies. Depending on the study design(memory clinic or population based) therate of conversion is between 10% and15% per year. MCI can also remain stable(as when due to stroke) or can resolveentirely (when due predominantly todepression). The operational definition ofMCI appears to be broadening beyonda purely amnestic entity. In a report ofthe International Working Group on MCI,the definition was revised to include thefollowing: (i) the person is neither normalnor demented; (ii) there is evidenceof cognitive deterioration measuredobjectively over time and/or a subjectivereport of decline by self and/or informantin conjunction with objective cognitivedeficits; and (iii) preservation of activitiesof daily living. Indeed multiple domain-MCI appears to be more prevalentthan pure amnestic-MCI in communitybased studies. A strict definition of MCIas a pure amnestic entity may be anoversimplification and not in line withday-to-day clinical practice where deficitsare not confined to memory loss but mayinvolve subtle cognitive impairmentsin the workplace or in the home. Inline with this revised approach, we areusing a broader definition of MCI andsub-classifying subjects into amnestic-MCI, multiple domain-MCI and singlenon-memory domain-MCI (eg in whichlanguage, visuospatial or executivefunction is impaired in isolation). Our MCIcohort includes a greater proportion ofsubjects that will subsequently convertto non-AD forms of dementia (multipledomain-MCI with and without amnesticfeatures and single non-memory domain-MCI). The clinical challenge is to predictthe ultimate pathological-diagnosis at thetime of presentation with reported earlycognitive impairment regardless of thecognitive domain(s) affected. Our studyAIM is to test a diagnostic algorithm thatincludes basic clinical and psychometricassessment and a combination ofimaging (MRI, FDG-PET, PIB-PET andVMAT PET), blood and cerebrospinalfluid (CSF) biomarkers on patientspresenting with a range of defined MCIsubtypes. CSF has been collected inmany patients and detailed imaging bothPIB-PET and VMAT PET undertaken on asignificant cohort of patients. VMAT-PETresults are robust and correlate with theclinical Parkinsonian phenotype in allpatients (PD and DLB) examined to date.The data validating the VMAT ligandhas been published. The preliminarydata suggests that it be possible todifferentiate between subgroups of MCIat presentation.Staff ListFellows and Research OfficersJohn Drago MB BS PhD (Melb)FRACP NHMRC Practitioner FellowResearch AssistantsJim Massalas Dip Lab TechStudentsLuning Jiang BSc (Hons)Hyun Ah Kim BSc (Hons)Postdoctoral FellowsShohreh Majd BSc PhDArthur Smardencas BSc PhDMajor Collaborative LinksNationalProf Andrew LawrenceFlorey Neuroscience InstitutesProf Colin MastersUniversity of MelbourneBiomarkers in MCIProf Chris RoweAustin HealthPET biomarkers in neurodegenerativediseasesDr Victor VillemagneAustin HealthPET biomarkers in neurodegenerativediseasesInternationalProf John WaddingtonRoyal College of Surgeons, IrelandMechanism of dystonia in ADNFLEmodels and cell ablation modelsof Huntington’s disease. Molecularmechanisms of D1 epileptogenesis9

Behavioural Neuroscience— Molecular NeurobiologyProf Ann Graybiel, MIT, Boston, USADepartment of Brain and CognitiveSciences, Building E25, Room 618,MIT,Cambridge, MA 02139,USAStudies to identify the anatomicalsubstratum of dystonia and Parkinsonism;the value of regional specific D1dopamine receptor positive cell ablationmice AND disruption of the striosomematrix architecture in D1 dopaminereceptor-expressing cell ablation mutantsProf Satoshi GotoDepartment of Neurosurgery,Kumamoto University Medical School,Kumamoto, JapanStudies to identify the anatomicalsubstratum of dystonia and Parkinsonism;the value of regional specific D1dopamine receptor positive cell ablationmice AND disruption of the striosomematrix architecture in D1 dopaminereceptor-expressing cell ablation mutantsVisiting ScientistsProf Katsunori TomiyamaDepartment of Pharmacology,Nihon University School of Dentistry,Tokyo, JapanPublicationsCannata D.J., Finkelstein D.I., Gantois I.,Teper Y., Drago J., West J.M.Altered fast- and slow-twitch musclefibre characteristics in female mice witha (S248F) knock-in mutation of the brainneuronal nicotinic acetylcholine receptor.J Muscle Res Cell Motil. 2009; 30(1-2):73-83.Fernando C.V., Parish C., Moses D.,Tomas D., Drago J., Horne M.K. Creatinga ventral midbrain stem cell niche in ananimal model for Parkinson’s disease.Stem Cells Dev. 2010 Apr 23. [Epubahead of print] PMID: 20415529.Okamura N., Villemagne V.L., DragoJ., Pejoska S., Dhamija R.K., MulliganR.S., Ellis J.R., Ackermann U., O’KeefeG., Jones G., Kung H.F., PontecorvoM.J., Skovronsky D., Rowe C.C. In vivomeasurement of vesicular monoaminetransporter type 2 density in Parkinsondisease with (18)F-AV-133. J Nucl Med.2010 Feb;51(2):223-8. Epub 2010 Jan15.PMID: 20080893 [PubMed – indexedfor MEDLINE]Babovic D., Jiang L., Gantois I., LawrenceA.J., Ferreri V., Schütz G., WaddingtonJ., Drago L. Age-related behaviouralphenotype and cellular characterisationof mice with progressive ablation of D1dopamine receptor-expressing cells. J.Behav Brain Res. 2010 Jan 5;206(1):78-87. Epub 2009 Sep 4.PMID: 19733597[PubMed - indexed for MEDLINE]10

Behavioural Neuroscience— Addiction NeuroscienceResearch OverviewOur laboratory is investigating theneural pathways implicated in drugseekingbehaviour and additionally, theassociation between mood disorders,stress and drug-seeking behaviour.For example, alcohol is one of themost widely used and abused drugsin society, with an immense social,medical and financial impact. Theneuropharmacological basis of alcoholreward is an ongoing research area inthis laboratory; we employ a number ofgenetic animal models, including inbredstrains of rats and knockout / transgenicmice. Drug self-administration andbehavioural parameters are measured toassess the influence of specific receptorsand emotional states such as anxiety anddepression on drug-seeking behaviour.Stressors can be used to examine therelationship between psychological stateand relapse to drug-seeking behaviourfollowing a period of abstinence.These behavioural approaches arecomplemented with a range ofneurochemical, anatomical andmolecular strategies to provide amultidisciplinary strategy.In addition, we are employing geneticapproaches to investigate the neuralsubstrates of drug-seeking, drug-inducedplasticity and relapse. This latter aspectis of critical importance, as the definingfeature of addictions is the chronic andrelapsing nature of the disorder. Manyof these research directions have acommon underlying theme of neuralplasticity as a key mediator of alteredbehaviour patterns. To facilitate thesestudies we have perfected the techniqueof intravenous self-administration ofdrugs of abuse in mice. Projects currentlyunderway involve self-administration ofopiates, cocaine and nicotine.Major Research ProjectsNeuropeptide involvement inalcohol-seekingAndrew Lawrence, Bianca Jupp,Michael Cowen, Betty Krivdic andElena KrstewCRF and orexin are neuropeptidesimplicated in a range of functionsincluding arousal, stress responses anddrug-seeking. We, and others, havepreviously demonstrated that bothCRF and orexin have roles in alcoholconsumption, seeking and relapse.Recently, we have developed a model ofrelapse to alcohol-seeking after extendedperiods of abstinence, demonstrating theutility of animals to study the enduringpropensity to relapse that is characteristicof human addicts. Importantly, wehave examined patterns of neuralactivation following cue-induced relapseimmediately after extinction training (arodent equivalent of rehab) comparedto relapse that occurs when extinctionis followed by an extended period ofabstinence. Immediate reinstatementincreased neural activation in a rangeof cortical and sub-cortical structuresimplicated in drug/alcohol-seeking andwhile delayed reinstatement showed abroadly similar pattern, these markerswere further elevated in cortical structures.These data suggest that a base networkof interconnected circuitry is implicated inrelapse-like behaviour, although followingprolonged abstinence the way the brainintegrates the salience of cues thatpreviously signalled drug availability maychange. Notably, functional blockade ofthe orexin system essentially preventedthe expression of relapse-like behaviourat both time points. This would suggestthat the orexin system in the brain isinvolved in cue-driven reward-seeking,irrespective of a period of abstinence.By combining pharmacological blockadeof the orexin system with the relapseparadigm and subsequent assessmentof neural activation we have identifiedputative anatomic loci where the orexinsystem may be acting to influencereward-seeking. These target structureswill be the focus of future studies toidentify where in the brain this peptidesystem acts to modulate relapse.Glutamate – adenosine interactionsand drug-seekingAndrew Lawrence, Robyn Brown,Michael Bird, Cameron Adams,Jennifer Short, Peter Dodd andKevin PflegerThis project has evolved out of ongoingstudies that have for some time beenexamining independently the role ofglutamate and adenosine systems indrug-seeking. For example, using acombination of pharmacological andgenetic approaches, we have previouslylinked the mGlu5 receptor to alcoholreward, motivation and sensitivity inboth rats and mice. More recently, usingmice with the adenosine A2a receptordeleted, we have linked this receptorto the reinforcing and motivationalproperties of opiates. For example,mice lacking the adenosine A2a receptorshow reduced desire to intravenouslyself-administer opiates compared towildtype littermates. Notably, alcoholand opiates act on similar pathways inthe brain; this is why naltrexone (anopioid receptor antagonist) can beused to reduce alcohol consumption.We therefore have provided a clear basefor the involvement of glutamate andadenosine in drug-seeking behaviour.Given that mGlu5 and adenosine A2areceptors are co-localised on a discretepopulation of neurons implicated in theneural network that is targeted by drugsof addiction, we hypothesised that thesereceptors may interact in a functionalmanner to regulate goal-directedbehaviour such as self-administration ofa drug. To test this, we trained a groupof rats to self-administer alcohol andthen injected them with a combinationof mGlu5 and adenosine A2a receptorantagonists. Importantly, we used dosesof antagonists that we had previouslydetermined were without effect bythemselves (ie subthreshold). When wecombined subthreshold doses of thetwo antagonists there was a dramatic,synergistic effect whereby alcohol selfadministrationwas reduced by almost50%. In addition, this combination ofmGlu5 and adenosine A2a receptorantagonists completely blocked relapseto alcohol-seeking in rats that had beenthrough a rehabilitation program.11

Behavioural Neuroscience— Addiction NeuroscienceThis project is now moving into humanbrain to study the interactions betweenthese two receptors and we are alsoscreening compound libraries forpotential lead compounds that mayinteract with these two receptors.Other Research ProjectsModelling inhalant abuse in rodentsAndrew Lawrence, Jhodie Duncanand Alec DickThe purposeful abuse of inhaled chemicalvapors to produce self-intoxication and/oraltered mental state is a significant publichealth concern. In Australia, sniffing thefumes of aerosol spray paint is the mostpopular form of inhalant abuse, whilepetrol sniffing remains a significantproblem for indigenous communities,especially within remote settings. Thetypical onset of experimentation withinhalants occurs earlier than with mostother drugs of abuse, in the preteen years,coinciding with the maturation of crucialcognitive and emotional brain structures.Despite long-standing awareness ofthe significant morbidity and mortalityassociated with inhalant abuse,fundamental neurobiological research hasbeen comparatively sparse. Accordingly,we have established a paradigmwhereby adolescent rodents inhalea major psychoactive component ofmany inhalants. Longitudinal studies arecurrently underway, incorporating stateof-theart Magnetic Resonance Imagingtechniques, to ascertain the impact ofchronic intermittent exposure to inhalantson brain pathology, in particular whitematter tracts. Parallel cohorts of rodentsare being examined to characterise theimpact of inhalant abuse upon affect,cognition and drug use in adulthood.The role of CREB and relatedtranscription factors in drug-seekingbehaviour and drug related plasticityAndrew Lawrence, CameronMcPherson, Heather Madsenand Srigala NavaratnarajahThe transcription factor CREB has beenimplicated in the consolidation of neuralplasticity associated with learning,memory and also drug addiction. Toaddress the latter possibility, we haveproduced lines of double transgenicmice where the CREB gene has beendeleted from defined populations ofneurons. Examination of one of theselines, where CREB has been deletedfrom glutamatergic telencephalicneurons suggests while that CREB inthese neurons is not involved in cocaineinduced plasticity, it is apparentlyinvolved in the motivation to selfadministercocaine. Parallel studieshave shown that CREB signals througha molecule called CREB binding protein(CBP) in medium spiny neurons of thebasal ganglia to regulate the behaviouralsensitivity to psychostimulants includingamphetamine and cocaine. Importantly,our basal ganglia model has the addedfeature that the genetic deletion of CREBor CBP occurs when mice are ~5 weeksold, preventing the occurrenceof adaptive compensation duringembryonic development.Nicotinic receptors and nicotine abuseAndrew Lawrence, John Drago,Liz Cahir and Katie PillidgeNicotine addiction, through smokingcigarettes, is a major contributor to theglobal burden of disease and a leadingcause of preventable death. To addressthis issue we have established a model ofnicotine self-administration in mice andare combining this with molecular geneticapproaches to examine the role of specificnicotinic receptor subunits in nicotinereward, plasticity, locomotor behaviourand anxiety. Ongoing experiments haveimplicated individual nicotinic receptorsubunits as necessary for differentbehavioural effects of nicotine.Additional ProjectsAnimal models of adolescent drug useAndrew Lawrence, Travis Featherby,Cameron McPherson and Elena KrstewGenetic models of drug-seekingbehaviourAndrew Lawrence, Bianca Jupp, JenniferShort, Robyn Brown, Michael Bird,Cameron McPherson, Heather Madsen,Liz Cahir, Katie Pillidge and Billy WestPublicationsAdams, C.L., Cowen, M.S., Short, J.L.& Lawrence, A.J. (2008) Combinedantagonism of glutamate mGlu5 andadenosine A2A receptors interact toregulate alcohol-seeking in rats. Int. J.Neuropsychopharmacol., 11, 229-241.Martin,S., Markus, M.A., Morris, B.J.,Davisson, R.L., Lawrence, A.J. & vanden Buuse, M. (2008) Does angiotensininteract with dopaminergic mechanismsin the brain to modulate prepulseinhibition in mice? Neuropharmacology,54, 399-404.Bird, M.K., Kircchoff, J., Djouma, E. &Lawrence, A.J. (2008) Metabotropicglutamate 5 receptors regulatesensitivity to ethanol in mice. Int. J.Neuropsychopharmacol., 11, 765-774.Featherby, T., van den Buuse, M., Lubman,D.I. & Lawrence, A.J. (2008) Persistentdown regulation in hippocampal CREBmRNA parallels a Y-maze deficit inadolescent rats following semi-chronicamphetamine administration. Br. J.Pharmacol., 154, 417-428.Lee, J., Zhu, W-M., Stanic, D., Finkelstein,D.I., Horne, M.H., Henderson, J.,Lawrence, A.J., O’Connor, L., Tomas,D., Drago, J. & Horne, M.K. (2008)Sprouting of dopamine terminals andaltered dopamine release and uptakein Parkinsonian dyskinesia. Brain, 131,1574-1587.Horne, M.K., Lee, J., Chen, F., Lanning,K., Thomas, D. & Lawrence, A.J. (2008)Long term administration of cocaine orserotonin reuptake inhibitors results inanatomical and neurochemical changesin noradrenergic, dopaminergic andserotonin pathways. J. Neurochem., 106,1731-1744.Lawrence, A.J. (2008) The demon drink:what’s on the horizon? Chemistry inAustralia, 75, 16-18.Lubman, D.I., Yücel, M. & Lawrence, A.J.(2008) Inhalant abuse among adolescents:neurobiological considerations. Br. J.Pharmacol., 154, 316-326.Brown, R.M., Short, J.L., Cowen, M.S.,Ledent, C. & Lawrence, A.J. (2009)A differential role for the adenosineA2A receptor in opiate reinforcementversus opiate-seeking behavior.Neuropsychopharmacology, 34, 844-856.12

Arolfo, M.P.*, Overstreet, D.H.*, Yao, L.,Fan, P., Lawrence, A.J., Tao, G., Keung,W.M., Vallee, B.L., Olive, M.F., Gass, J.T.,Rubin, E., Anni, H., Hodge, C.W., Besheer,J., Zablocki, J., Leung, K., Blackburn,B.K., Lange, L.G. & Diamond, I. (2009)Suppression of heavy drinking andalcohol-seeking by a selective ALDH-2inhibitor. Alc. Clin. Exp. Res., 33, 1935-1944.Bird, M.K. & Lawrence, A.J. (2009) GroupI metabotropic glutamate receptors:involvement in drug-seeking & druginducedplasticity. Curr Mol. Pharmacol.,2, 83-94.Brown, R.M. & Lawrence, A.J. (2009)Neurochemistry underlying relapse toopiate seeking behaviour. Neurochem.Res., 34, 1876-1887.Bird, M.K. & Lawrence, A.J. (2009) Thepromiscuous mGlu5 receptor – a rangeof partners for therapeutic possibilities?Trends Pharmacol. Sci., 30, 617-623.Lawrence, A.J. (2009) Factors regulatingstress-induced alcohol-seeking andpharmacotherapeutic treatments.Alcohol, 43, 545-546.13

Brain Development and RegenerationResearch OverviewMany adult brain disorders havetheir genesis during the assembly ofbrain cells in the fetus. Therefore, theunderstanding of how new brain cells aregenerated and connected together ispivotal to preventing and treating thesedisorders. The underlying drivers of braincell assembly are undoubtedly genetic,but the outcomes are also shaped byenvironmental influences. Our laboratoryhas been studying the roles of Reelin,Sez-6, and Ndfip1. Separately, each ofthese genes is important for controllingneuronal proliferation, neuronal migrationand dendritic maturation. Reelin isimportant for governing the way newneurons migrate to their destinationsand are organized. Sez-6 is important forcontrolling the branching of neurons andconnecting them together. Ndfip1 has apivotal role in influencing the generationof new neurons. Together, these studiesaccrue fundamental and detailedinformation behind the assembly ofbrain cells in the developing embryo.While the birth and maturation ofbrain cells are important for properneurological function, their survivaland protection against noxious stimuliare equally vital for the preservationof neuron number and brain function.For example, perinatal hypoxia poses asignificant risk to neuron survival duringbirth. Our work with brain injury modelshave identified an intrinsic defensemechanism that protects brain cellsfrom death following injury from hypoxia,trauma and ischemia. Our discovery thatNdfip1 is neuroprotective in brain injury isnow replicated in stroke, suggesting thatneuroprotection by Ndfip1 is a genericand conserve mechanism for defendingthe brain. This has spurred our researchactivity to higher levels, focussing onbiochemical and molecular mechanismsthat underscore Ndfip1 function. Ourover-riding aim is to discover howNdfip1 activity can be controlled bypharmacological means with the view oftherapeutic benefit to future sufferers ofbrain injury from trauma and stroke.Members of the Team have also engagedin strong collaborative activity leadingto published works in the field of neurontranscription (Hamish Scott), cocaineaddiction (Andrew Lawrence), Alzheimer’sDisease (Colin Masters), epilepsy(Steve Petrou) and retina development(Michael Kalloniatis).Major Research ProjectsHow interneurons find their wayduring migration?Joanne Britto, Leigh Johnstonand Seong-Seng TanDuring cortical development,interneurons migrate long distance toreach their destinations. In this study, weset out to investigate how interneuronsmigrate in real time, and attempt toestablish migration rules using computermodelling. Migration is a dynamicprocess in which a cell searches theenvironment and translates acquiredinformation into somal advancement.In particular, exploration in interneuronmigration during development isaccomplished by the extension ofneurites tipped with growth cones,whilst locomotion is achieved by nucleustranslocation, termed nucleokinesis.The primary purpose of our work wasto investigate the relationship betweenneurite branching and nucleokinesisusing high resolution time-lapseconfocal microscopy and computationalmodelling. We found that nucleokinesisis accurately modelled by a criticallydamped mass-spring-damper systemand that neurite branching, with thedynamics of a stochastic birth-deathprocess, can be considered in isolation tothe event of nucleokinesis. This is contraryto traditional descriptive analyses thatassume a closer relationship betweenthe two migratory mechanisms. Ourmodels are validated on independentdata sets acquired using two differentimaging protocols, and are shown tobe robust to alterations in guidancecues and cellular migratory mechanisms,through treatment with Brain DerivedNeurotrophic Factor, neurotrophin-4,and blebbistatin. We postulate that thestochastic branch dynamics constitutean analogue Markov Chain MonteCarlo algorithm, used by interneuronsundergoing guidance directed migrationto efficiently explore the environment.Ndfip1 protects human neuronsfrom death by ubiquitination anddegradation of metal transporterDMT1Jason Howitt, Ulrich Putz, Jenny Lackovic,Anh Doan and Seong-Seng TanDuring brain injury, the homeostasis oftransition metal ions (eg Fe2+, Co2+)is grossly unbalanced. This rendersthe brain environment toxic due tounwelcomed and uncontrolled metalentry into stressed neurons, precipitatingtheir premature death. Using culturedprimary human neurons and alsoestablished cell lines, we demonstratethat Ndfip1 protects neurons againstexcess transition metal exposure. In thepresence of excess Co2+ or Fe2+ ions,Ndfip1 is upregulated. We discoveredthat the mechanism for this protection ismediated by the metal transporter DMT1.Ndfip1 binds to DMT1, leading to DMT1ubiquitination and degradation. In thismanner, metal entry is inhibited, allowingneuronal recovery. To test this idea invivo, we generated a mouse knock out forNdfip1. When fed with excess free Fe2+,these mice accumulate excess metal intheir brains. More recently, have exploredthe connection between Ndfip1 and braindiseases (eg Parkinsons Disease) arisingfrom metal accumulation.Seizure-Related Gene 6 (Sez-6) inAmacrine Cells of the Rodent Retinaand the Consequence of Gene DeletionJenny Gunnersen, Annabel Kuek,Joanna Phipps, Vicki Hammond,Theresa Puthussery, Erica Fletcherand Seong-Seng TanThe distribution pattern of Sez-6in the retina was studied using apolyclonal antibody that detects themultiple isoforms of Sez-6. Prominentimmunostaining was detected inGABAergic, but not in AII glycinergic,amacrine cell subpopulations of the ratand mouse retina. Amacrine cell somatadisplayed a distinct staining pattern withthe Sez-6 antibody: a discrete, oftenroughly triangular-shaped bright spotpositioned between the nucleus andthe apical dendrite superimposed overweaker general cytoplasmic staining.Displaced amacrines in the ganglioncell layer were also positive for Sez-6and weaker staining was occasionallyobserved in neurons with the morphology14

of alpha ganglion cells. Two distinctSez-6 positive strata were present inthe inner plexiform layer in addition togeneralized punctate staining. Certaininner nuclear layer cells, including bipolarcells, stained more weakly and diffuselythan amacrine cells, although somebipolar cells exhibited a perinuclear“bright spot” similar to amacrine cells.In order to assess the role of Sez-6 inthe retina, we analyzed the morphologyof the Sez-6 knockout mouse retinawith immunohistochemical markers andcompared ganglion cell dendritic arborpatterning in Sez-6 null retinae withcontrols. The functional importance ofSez-6 was assessed by dark-adaptedpaired-flash electroretinography (ERG).In summary, we have reported thedetailed expression pattern of a novelretinal marker with broad cell specificity,useful for retinal characterization in rodentexperimental models. Retinal morphology,ganglion cell dendritic branching andERG waveforms appeared normal in theSez-6 knockout mouse suggesting that,in spite of widespread expression ofSez-6, retinal function in the absenceof Sez-6 is not affected.Staff ListFellows, Senior Research Officersand Research OfficersSeong-Seng Tan BDS (Malaya) MDS (Adel),DPHIL(Oxon) FRACDSVicki Hammond BSc (Hons) BA PhD (Melb)Jenny Gunnersen BSc (Hons) (JCU) PhD (Melb)(Fractional appointment)Julian Heng BSc (Hons) (UWA) PhD (Melb)Joanne Britto BSc (Hons) PhD (Cantab)Jason Howitt PhD (Swinburne)Ulrich Putz PhD (Hamburg, Germany)Christine Scieber PhD (Melb)Research AssistantsEan-Phing LeeAhn DoanAlison McIntyreTomoko HyakumuraJoanne BarwoodNaomi CookeKaren TaitZheng-Dong QuStudentsChoo-Peng Goh BSc (Hons) (Monash)Jenny Lackovic BCom BSc (Hons) (Melb)(graduated 09/2010)Ley-Hian Low BSc (Hons) (Monash)Awards and ConferencePresentationsSeong-Seng TanJapan Neuroscience Society 31stAnnual Meeting Tokyo July 2008.Symposium “Regulation of NeuronalSurvival and Production in AdultBrain” Title: “Molecular mechanisms ofneuroprotection by Ndfip1 followingbrain injury”Keio Global COE InternationalSymposium “Mechanisms of CerebralCortical Development”, Keio, Japan July2008. Title: “Mechanisms steering themigration of cortical neurons”Tohoku University Symposium (Host ProfNoriko Osumi) Sendai, Japan July 2008.“Neuroprotection in brain injury andstroke – the role of protein ubiquitinationand exosomes”Awaji meeting on “Construction andreconstruction of the brain” Awaji IslandOctober 2009. “Protein ubiquitination byNdfip1 in cortical development and adultcortical function”RIKEN Center for Developmental Biology(Host Prof Fumio Matsuzaki) KobeDecember 2009. “How can a proteinthat protects the brain during injury beharmful during brain development?”Australian Neuroscience Society annualmeeting, Hobart, Jan 2008. OralPresentation “Ndfip1 neuroprotectionfollowing brain injury”Australian Neuroscience Societyannual meeting, Canberra, Jan 2009.Oral Presentation “Mechanisms ofinterneuron migration”Eccles Neuroscience Institute, ANUseminar series, Canberra, August 2009.“Cellular and molecular mechanisms ofneuroprotection by Ndfip1 in the brainfollowing trauma and stroke”Prince of Wales Medical ResearchInstitute, UNSW, Sydney July 2009.“Neuroprotection of the brain by Ndfip1”Queensland Brain Institute PublicLectures on Building a Brain, Brisbane,July 2009. “Building a Brain – harmminimization following injury”Monash University NeuroscienceSeminar Series, August 2009. “How cana molecule that protects the brain duringinjury be harmful during development”?Flinders University Medical CenterSeminar Series, November 2009.“How can a molecule that protectsthe brain during injury be harmfulduring development”?Joanne Britto2009 Australian Neuroscience Society29th Annual Meeting, Canberra AUSJason Howitt2nd Annual Melbourne Cell andDevelopmental Biology Meeting,Nov 2009. The regulation of DMT1by Ndfip1 prevents metal toxicitywithin neuronsJulian HengHeng, J.I., Qu, Z.D., Guillemot, F. andTan, S.-S. Negative feedback regulationof a proneural bHLH program forneurodifferentiation by a zinc fingertranscription factor. 32nd annual meetingof the Japanese Neuroscience Society,Nagoya (Japan), September 2009.Recipient of a JNS Travel Award forthis meetingHeng, J.I., Transcriptional Control ofNeuronal Migration in the DevelopingCerebral Cortex. 29th Meeting ofthe Australian Neuroscience Society.Canberra, Australia. January 2009.Heng, J.I., Transcriptional Control ofNeuronal Migration in the DevelopingCerebral Cortex. Inaugural MelbourneCell and Developmental Biology Meeting(VSDB, MEG, ANZSCDB), Melbourne.November 2008. Recipient of prize forTop Oral Presentation at this meeting.15

Brain Development and RegenerationVicki Hammond“Reelin signalling and neuronal migrationin the developing brain”. School ofPharmacy and Applied Science, La TrobeUniversity, Bendigo, (October 2009).Jenny GunnersenYoung Investigator Award 2009,International Society for Neurochemistry/Asia-Pacific Society for Neurochemistry22nd Biennial Joint Meeting, August 2009,Busan Korea.“How do neurons acquire theircharacteristic dendritic arbors?”Oral presentation in Young InvestigatorColloquium 7, entitled “Mechanisms ofPlasticity and Memory II”.Major Collaborative LinksNationalProf Perry BartlettQueensland Brain Institute,University of QueenslandGeneration of neurons in theadult and embryoProf Pankaj SahQueensland Brain Institute,University of QueenslandElectrophysiology of brain cellslacking Sez-6Prof Sharad KumarHanson Centre, IMVS, AdelaideThe role of Ndfip1 in brain injuryProf David VauxDepartment of Biochemistry,La Trobe UniversityNeuroprotective functions of Ndfip1Dr John SilkeDepartment of Biochemistry,La Trobe UniversityBiochemistry and molecular biologyof Ndfip1A/Prof Cristina Morganti-KossmannNational Trauma Research Institute,Alfred Hospital and Monash UniversityInternationalDr Zoltan MolnarUniversity of Oxford, EnglandThalamocortical innervation ofmutant cortexProf Baoli YangDepartment of Obstetrics andGynecology, Carver College of Medicine,Iowa City, USAThe role of Ndfip1 gene in braindevelopmentStudent graduatedPhD – Michaella Richards Title “The roleof the basic helix-loop-helix transcriptionfactor Bhlhb5 in cell fate determinationand the developing cortex”Visiting ScientistsProf Michael KalloniatisUniversity of Auckland, New ZealandEditorial PositionsSeong-Seng TanJournal of Neuroscience (USA)Experimental NeurologyJournal of AnatomyAnatomy & Cell Biology (Korea)PublicationsAcosta, M., Bumsted-O’Brien, K.,Tan, S-S., Kalloniatis, M. (2008).Emergence of cellular markers andfunctional ionotropic glutamate receptorson tangentially dispersed cells in thedeveloping mouse retina. J. Comp.Neurol. 506:506-523Voss, A.K., Britto, J.M., Dixon, M.P.,Sheikh, B.N., Collin, C., Tan, S-S., Thomas,T. (2008) C3G regulates cortical neuronmigration, preplate splitting and radialglial cell attachment. Development135:2139-2149Chiu, C., Reid, C.A., Tan, H.O., Davies,P.J., Single, F.N., Koukoulas, I., Berkovic,S.F., Tan, S-S., Sprengel, R., Jones, M.V.,Petrou, S. (2008). Developmental impactof a familial GABA A receptor epilepsymutation. Annals Neurol. 64:284-293Foot, N.J., Dalton, H.E., Shearwin-Whyatt, L., Dorstyn, L., Tan, S-S., Yang,B., and Kumar, S. (2008). Regulation ofthe divalent metal ion transporter DMT1and iron homeostatis by a ubiquitindependentmechanism involving Ndfipsand WWP2. Blood 112: 4268-4275Putz, U., Howitt, J., Lackovic, J., Foot,N., Kumar, S., Silke, J. and Tan, S-S.(2008). Nedd4-family interacting protein1 (Ndfip1) is required for the exosomalsecretion of Nedd4-family proteins.J. Biol. Chem 283:32621-32627Tan, S-S., Truong, H., Binder, M., Cate,H., Kilpatrick, T., Kalloniatis, M. andHammond, V. (2009). Oligodendrocytepositioning in cerebral cortex isindependent of projection neuronlayering. Glia 57:1024-1030Britto, J.M., Johnston, L.A. and Tan, S-S.(2009). The stochastic search dynamics ofinterneuron migration. The Biophysical J.97: 699-709Ling, K.H., Hewitt, C.A., Beissbarth,T., Hyde, L., Banerjee, K., Cheah, P.S.,Cannon, P.Z., Hahn, C.N., Thomas, P.Q.,Smyth, G.K., Tan, S-S., Thomas, T., Scott,H.S. (2009). Molecular networks involvedin mouse cerebral corticogenesis andspatio-temporal regulation of Sox4and Sox11 novel antisense transcriptsrevealed by transcriptome profiling.Genome Biol. 10(10):R104Gunnersen, J.M., Kuek, A., Phipps, J.A.,Hammond, V.E., Puthussery, T. , Fletcher,E. and Tan, S-S (2009). Seizure-RelatedGene 6 (Sez-6) in amacrine cells of therodent Retina and the consequence ofgene deletion. PLoS ONE 4(8):e6546Howitt, J., Putz, U., Lackovic, J., Doan, A.,Dorstyn, L., Cheng, H., Yang, B., Chan-Ling, T., Silke, J., Kumar, S. and Tan, S-S.(2009). Divalent metal transporter 1(DMT1) regulation by Ndfip1 preventsmetal toxicity in human neurons. Proc.Natl. Acad. Sci. USA 106:15489-15494Xiao, J., Wong, A.W., Willingham, M.M.,Kaasinen, S.K., Hendry, I.A., Howitt, J.,Putz, U., Barrett, G.L., Kilpatrick, T.J. andMurray, S.S. (2009). J Neurosci. 29:4016-4022.16

Heng, JI, Chariot, A and Nguyen, L.Molecular layers underlying cytoskeletalremodelling during cortical development.Trends in Neurosciences. 2010Jan;33(1):38-47Merot, Y, Reteaux, S and Heng, JI.Molecular mechanisms of projectionneuron migration and maturation in thedeveloping cerebral cortex. Seminars inCell and Developmental Biology. 2009Aug;20(6):726-34.Geng G, Johnston LA, Yan E, Britto JM,Smith DW, Walker DW and Egan GF(2009) Biomechanisms for modellingcerebral cortical folding. Medical ImageAnalysis 13:920-93017

Clinical Trials andNeuroscience Trials AustraliaOverviewThe clinical trials division continues toexpand with about thirty trials beingconducted at any one time. These consistof investigator initiated studies as well asthose initiated by commercial interests.Investigator driven trials such as ARCH inwhich patients in whom the mechanism ofischaemic TIA or stroke is most likely dueto the presence of aortic arch atheromaare randomised to receive eithercombination antiplatelet therapy orthe more traditional anticoagulant,warfarin continue.Our other main interest continues to bein acute stroke therapy (neuroprotectionand thrombolysis) as well as secondaryprevention. As part of our approach tothe former, the neuroimaging modalitiesare increasingly being used (particularlyMRI) to provide important “proof ofconcept” information before embarkingupon larger clinical trials. The phase IIMR based trial, EPITHET, co-chaired byProfessor Stephen Davis and GeoffreyDonnan has been completed and hasnow been published in the prestigiousLancet Neurology Journal. The findings ofa significant increase in cerebral perfusionparameters provided strong biologicalsupport for the idea that the time windowfor thrombolysis may be extended from3 to 6 hours. A new trial with MR basedimaging selection criteria (EXTEND) willcommence during 2009.A major focus of this program is A VeryEarly Rehabilitation Trial (AVERT) whichis now in Phase III. In this innovativestudy, the hypothesis that earlier andmore intensive mobilisation of strokepatients will improve outcomes, and becost effective, is being tested (n=2104).In 2009, we expanded the number ofrecruiting centres to 38, and commencedrecruitment in Singapore and Malaysia.Recruitment sits at 745 patients andAVERT is already the largest strokerehabilitation trial in the world. In 2009we were successful in obtaining a StrokeAssociation UK grant to further expandAVERT. This will take place throughout2010. Associate Professor Julie Bernhardtis Director of the Very Early RehabilitationResearch Program which is focused notonly on efficacy and cost of very earlyinterventions but also mechanismsof action of these interventions andimplementation research.In the secondary prevention field, a largenumber of studies are continuing to beconducted using antiplatelet agentsand anticoagulants as well as agentsdesigned to reduce the risk ofsurgical procedures such as carotidendarterectomy. The latter trial, Dextranin Carotid Endarterectomy (DICE) is oneof the largest of its type ever conductedand is now nearing completion.Any clinical trial is highly dependentupon collaborative links with a largenumber of centres both within Australiaand internationally. In this regard, weare particularly fortunate in having theAustralian Stroke Trials Network (ASTN)and well established internationallinkages. A further initiative is theformation of Neuroscience Trials Australia(NTA) which Professor Geoff Donnan cochairswith Professor Stephen Davis at theRoyal Melbourne Hospital. Here, a seriesof nodes from almost every neurosciencegroup (such as stroke, epilepsy, migraine,movement disorders etc) form a singleorganisation to attract more trials toAustralia as well as providing supportfor investigator driven studies. The NTAManager, Dr Peter Keller, has alreadybeen successful in attracting newinitiatives on to the Australian scene, anexcellent sign for the future.Major Research ProjectsAVERT (A Very Early Rehabilitation Trial)Julie Bernhardt, Helen Dewey,Geoffrey Donnan, Amanda Thrift,Janice Collier, Richard Lindley, MarjMoodie, Peter Langhorne and theAVERT Trialists CollaborationStroke presents a major, growingglobal public health challenge whichaccounts for 25% of all chronic disability.Treatments that reduce the personal andcommunity burden of stroke are urgentlyneeded, and the commencement ofrehabilitation early after stroke maysignificantly reduce chronic disability.A large (n=2104), high quality, NH&MRCfunded clinical trial is at the heart of theAVERT program. The trial tests whethera simple, easy to apply rehabilitationintervention (early and intensive out ofbed activity) results in fewer deaths, lessdisability and better long term quality oflife for stroke sufferers. In addition, wewill examine whether the intervention isalso cost effective. If effective (and costeffective) the intervention should bereadily adopted within a diverse rangeof health services both here and overseas,and help reduce the global burdenof stroke.The trial commenced in July 2006 at9 Australian hospitals, following onfrom successful Phase I and II studieswhich showed the need for an earlyrehabilitation intervention withinAustralian hospitals and that theintervention protocol developed forthe study was both safe and feasible.The inclusion criteria for this study arebroad with no upper age limit, firstor subsequent stroke, infarct (clot) orhaemorrhage (bleed). The trial design ispragmatic with all research staff involvedin this study part of a functioning strokeunit service. The trial intervention isdelivered by a nurse/physiotherapy teamand these clinicians are the drivers ofthe study at each site. In early 2008 wedetermined that the study needed to beexpanded across more sites to improverecruitment. We grew from 9 sites to nowover 38 sites with significant ongoinginternational expansion expected.Prof Peter Langhorne (Scotland, UK) andA/Prof Bernhardt and her team workedtogether to establish 5 Scottish sites in2009 and this paved the way for furthersuccess with the Stroke Association grant(GBP 186,000) at the end of 2009. Twosites in Wales were also trained in 2009and Drs Shiela Lennon and MichaelPower (Northern Ireland, UK) wereawarded $200,000 to support sites inNorthen Ireland, with Belfast and UlsterHospitals now recruiting. We expect trialrecruitment to close in 2012.Sophisticated trial software developedthrough a student partnership betweenRMIT University and the National StrokeResearch Institute supports the smoothrunning of this trial. The managementteam for this project is located at theHeidelberg Repatriation Hospital,Melbourne.EXTEND trialGeoff Donnan, Stephen Davis(Royal Melbourne Hospital CollaboratingCentre) and Helen DeweyThe EXTEND trial builds on the dataabout the safety and efficacy ofthrombolysis provided by EPITHET18

eyond the 3 hour time window. Inthis new phase III trial we plan toselect patients most likely to respondto thrombolysis based on the MR orCT imaging presence of significantslavageable (penumbral) brain tissue.The tiem window for thrombolysiswithin this randomized double blinded,controlled trial will be extended up to9 hours past stroke onset. Centresinvolved in the study will be in Australia,New Zealand, Asia and Europe. This willbe launched during 2010.Other Research ProjectsThe effect of early and intensiverehabilitation on mood and cognition(AVERT sub-study)Toby Cumming, Thomas Linden(Gothenberg, Sweden) and Julie BernhardtApproximately one third of strokesurvivors become depressed and upto two thirds experience cognitiveimpairment (eg, problems with payingattention, remembering information andcommunicating effectively). Not only aredepression and cognitive impairmentmajor problems in themselves, they areassociated with other negative outcomesincluding earlier death, greater rates ofinstitutionalisation, poorer quality of lifeand higher health care costs. Findingsimple, easy to apply interventionsthat protect against depression andcognitive impairment after stroke wouldbe extremely valuable in improvingoutcomes for stroke survivors.Increasing early physical activity afterstroke is a very attractive intervention:broadly applicable, inexpensive andwidely available. At present, there is noclear evidence that exercise early afterstroke reduces depression or cognitiveimpairment. The AVERT trial providesa unique and exciting opportunityto test this possibility. In 2008 wepublished results from Phase II AVERTdemonstrating that early and frequentmobilisation after stroke was associatedwith reduced depression at 7 days poststroke.Assessing cognitive function afterstroke is more difficult – no measure thatis brief, easy to administer and valid instroke has been identified. For this reason,we are running a study with the aim ofidentifying a valid and feasible screeningtool for cognitive impairment after stroke.Recruitment to this study is steady andwe expect to complete the study in 2011.Phase III AVERT includes depressionassessment at 3 and 12 months poststroke,and now also includes cognitiveassessment at 3 months. A cohort of morethan 300 stroke patients have been testedon this cognitive assessment to date, andthese results are currently being writtenup for publication. At the conclusionof the trial, We aim to determine theeffect of increased physical activity ondepression and cognition following stroke,and isolate the undeyling mechanisms.This will require the use of assessments ofmood and cognitive function, electronicactivity monitors to record continuousphysical activity, blood samples tomeasure relevant physiological markers,and brain imaging to detect structuraland functional changes brought aboutby exercise.DICEGeoff DonnanThe DICE trial is nearing completion.Here the hypothesis that the surgicalprocedure of carotid endarterectomywould result in few adverse stroke andTIA outcome events if patients receivedintravenous dextran perioperatively.The study design is that of a phase IIIrandomized double blinded controlledtrial with clinical outcome measures withinone month. Regardless of the results, thefindings will significantly influence clinicalmanagement world wide and will becompleted during 2010.Additional Research Projects(clinical trials)ARCH CLINICGeoff DonnanVITATOPSGeoff Donnan, Helen Dewey andGraeme Hankey (Royal Perth Hospital)ENOSHelen DeweyICSSBrian ChambersINTERACT 2Helen DeweyIST 3Helen DeweyBone and muscle loss after stroke andtheir relationship to physical activityJulie Bernhardt, Marco Pang (Hong KongUniversity), Sandy Luliano (EndocrineCentre of Excellence, Austin Health),Coralie English (University of SouthAustralia) and Karen BorschmannStaff and StudentsGeoffrey DonnanMBBS, MD, FRACP, FRCP (Edin)Dennis Young, Graduate Certificate of Nursing(Critical Care), Certificate of Nursing, AssociateDiploma of Welfare Studies, Diploma of ArtsLisa WalkerRoslyn Melling, Clinical Trials Co-ordinatorSandra Petrolo, Stroke Trials Co-ordinatorAVERTStaffJulie Bernhardt BSc PhDJanice Collier BSc, MA. PhDFiona Ellery B NursingJan Chamberlain BSc, Grad Dip Ed,Adv Cert HortToby Cumming BBSc (Hons) PhDNicholas Haritos BScCecilia Li BCom/BScHenry Zhao BScKaren Borschmann BScSally Speare BScStudentsLouise Craig BSc PhD Student (UK)Sharon Kramer BSc Masters Student(Netherlands)Karen Tyedin BSc Honours StudentGeraldine Banks BSc Honours StudentRenske van Wijk BSc Masters (Netherlands)Rosalie Kroeders BSc, Advanced MedicalScience Student (Netherlands)19

Clinical Trials andNeuroscience Trials AustraliaTara Purvis BSc Masters StudentRenee Trigg BSc Professional DoctorateStudentJohn Nguyen BSc Honours StudentJason Ha BSc, Advanced MedicalScience StudentKaren Borschmann BSc Masters StudentWenwen Zhang BSc PhD StudentJacinta Simek BSc Honours StudentHeidi Janssen B Sc PhD StudentCoralie English BSc, PhD, Post Doctoral FellowJulie Luker BSc, PhD StudentTanya West BSc, Masters StudentLeigh Wittle B App Sc. StudentAileen Ibuki BSc, PhD StudentAnna Sjoholm BSc Student (Netherlands)Monica Skarin BSc Student (Netherlands)Neuroscience Trials AustraliaPeter Keller BAppSC(Optom), MBA, PhD,MHEth, PGCertOcTher, FVCOAnita Thompson Administrative OfficerSue Bates BSc (Hons), GradCertHlthServMt[Graduate Certificate Health ServicesManagement]Fiona Ellery Bachelor of NursingAngela JamesJacqui HoustonNick TiliacosCara ThompsonConferences andPresentationsEvidence-based Management of AcuteIschaemic Stroke and Prevention ofRecurrent Events. 18th European StrokeConference, Stockholm, Sweden,May 26-29, 2009Acute Stroke Therapy & Prevention:“Stroke as an Emergency”, WorldCongress of Neurology, Bangkok,October 24, 2009“New Insights on Stroke Prevention”,World Congress of Neurology, Bangkok,October 24, 2009National and InternationalCollaborative LinksInternationalProf Pierre Amarenco, Paris, FranceARCHProf Ken Lees, Glasgow, UKVISTADr Malcolm Macleod, Edinburgh, UKARCHProf Greg Albers, Stanford StrokeCentre, Stanford University MedicalCentre, San Fransisco, USAEPITHETDr Steven Warach, NIH, Bethesda, USAMR Stroke CollaborationProf Werner Hacke, University ofHeidelberg, GermanytPA meta analysesProf P Langhorne, Glasgow RoyalInfirmary, Glasgow, UKStroke rehabilitation expertProf B Indredavik, St Olav’s Hospital,Trondheim, NorwayVery early rehabilitation expertM Halvorsen, Karolinska Institute,Stockholm, SwedenStroke unit evaluation projectProf van Meeteren, Utrecht University,Netherlands. Student exchangeprogramTranslation of evidence to practice instroke rehabilitationAssistant Prof M Pang, Hong KongPolytechnic University, Hong KongBone loss after strokeDr Thomas Linden, GotenborgUniveristy Hospital, SwedenInfluence of exercise on post strokedepression and cognitionAVERT Collaboration – Stroke Unit TrialistsCanada: Medicine HatMalaysia: UKMNew Zealand: AucklandSingapore: Singapore GeneralUnited Kingdom: Royal Glasgow;Wishaw General; Stirling Infirmary;Monklands, Crosshouse, Belfast City,Ulster. Royal Gwent, Nevill HallNationalAVERT Collaboration – Hospital StrokeUnit TrialistsNew South Wales: Belmont, Gosford,Newcastle Mater, St Vincent’s, Westmead,Wollongong, WyongQueensland: Nambour, WesleySouth Australia: Flinders Medical CentreVictoria: Austin, Frankston, Geelong,Royal Melbourne, St. Vincent’s, The Alfred,Warrigal, WesternWestern Australia: Sir Charles Gairdner,Royal PerthRMIT University, Melb. SoftwareEngineering Department.Developed web-based randomisationand data management platform for PhaseIII study to our specs.Prof L Carey, La Trobe University, MelbAssisted with development of tool foranalysis of hand motion.Prof R Carter, Deakin University, HealthEconomics DepartmentCost effectiveness evaluation.Dr S Luliano, Endocrine Centre ofExcellence, University of MelbourneRole of early rehabilitation on nutrition/lean body mass and glucose metabolism.Dr Neil Spratt, Newcastle University,Newcastle NSWSystematic reviews of animal literatureexamining enriched environment andexercise in stroke rat models.Editorial PositionsProf Geoffrey DonnanCerebrovascular DiseasesStroke20

CNS DrugsJournal of Clinical NeurosciencesJournal of NeuroimagingJournal of Internal MedicineLancet NeurologyInternational Journal of StrokeJournal of the CardioMetabolic SyndromeAnnals of Indian Academy of NeurologyA/Prof Julie BernhardtTopics in Stroke RehabilitationInternational Journal of Therapy andRehabilitationStroke Research and Treatment (GuestEditor)Visiting ScientistsDr Coralie English, South Australia,AustraliaDr Marco Pang, Hong Kong University,Hong KongDr Thomas Linden, SwedenMs. Louise Craig, Scotland, UKMs. Monica Skarin, SwedenMs. Anna Sjoholm, SwedenDr Torunn Askim, NorwayMs Renske van Wijk, The NetherlandsMs Sharon Kramer, The NetherlandsMs Roos Kroeders, The NetherlandsDr Sheila Lennon-Fraser, Northern IrelandMs Dawn Tan, SingaporeAwardsA/Prof Julie BernhardtAustralian Research Council Future FellowAwardDr Toby CummingStroke Society of Australasia overseastravelling fellowshipPublicationsAmarenco P, Bogousslavsky J, Caplan LR,Donnan GA, Hennerici MG. Classificationof Stroke Subtypes. Cerebrovasc Dis2009;27(5):493-501.Purvis T, Cadilhac D, Donnan G,Bernhardt J. Systematic review of processindicators: including early rehabilitationinterventions used to measure qualityof acute stroke care. Int J Stroke 2009Apr;4(2):72-80.Amarenco P, Bogousslavsky J, CaplanLR, Donnan GA, Hennerici MG. NewApproach to Stroke Subtyping: The A-S-C-O (Phenotypic) Classification of Stroke.Cerebrovasc Dis 2009;27(5):502-8.Bladin C, Chambers B, Crimmins D, DavisS, Donnan G, Frayne J, Levi C, DentonM, Lawrence-Brown M, Phatouros C,Scroop R, Harrington T, Carotid StentingGuidelines C. Guidelines for patientselection and performance of carotidartery stenting. J Med Imag Radiat Oncol2009 Dec;53(6):538-45.Macleod MR, Fisher M, O’Collins V,Sena ES, Dirnagl U, Bath PMW, BuchanA, van der Worp HB, Traystman R,Minematsu K, Donnan GA, Howells DW.Good Laboratory Practice PreventingIntroduction of Bias at the Bench. Stroke2009 Mar;40(3):E50-E2.Macleod MR, Fisher M, O’Collins V, SenaES, Dirnagl U, Bath PMW, Buchan A, vander Worp HB, Traystman RJ, MinematsuK, Donnan GA, Howells DW. Reprint:Good laboratory practice: preventingintroduction of bias at the bench (vol 29,pg 221, 2009). J Cereb Blood Flow Metab2009 Jun;29(6):1226-.Ninomiya T, Donnan G, Anderson N,Bladin C, Chambers B, Gordon G, SharpeN, Chalmers J, Woodward M, Neal B, GrpPC. Effects of the End Point AdjudicationProcess on the Results of the PerindoprilProtection Against Recurrent Stroke Study(PROGRESS). Stroke 2009 Jun;40(6):2111-5.Sorbello D, Dewey HM, Churilov L, ThriftAG, Collier JM, Donnan G, Bernhardt J.Very Early Mobilisation and Complicationsin the First 3 Months after Stroke: FurtherResults from Phase II of A Very EarlyRehabilitation Trial (AVERT). CerebrovascDis 2009;28(4):378-83.Thrift AG, Dewey HM, Sturm JW, SrikanthVK, Gilligan AK, Gall SL, MacdonellRAL, McNeil JJ, Donnan GA. Incidenceof Stroke Subtypes in the North EastMelbourne Stroke Incidence Study(NEMESIS): Differences between Menand Women. Neuroepidemiology2009;32(1):11-8.Venketasubramanian N, Chen CLH,Gan RN, Chan BPL, Chang HM, TanSB, Picard D, Navarro JC, Baroque AC,Poungvarin N, Donnan GA, Bousser MG,Investigators C. A double-blind, placebocontrolled,randomized, multicenterstudy to investigate CHInese MedicineNeuroaid Efficacy on Stroke recovery(CHIMES Study). Int J Stroke 2009Feb;4(1):54-60.Tyedin K, Cumming T, Bernhardt J. (Inpress). Quality of life. An important butcomplex measure in a trial of very earlymobilisation after stroke. Disability &Rehabilitation. 2009Wellwood I, Langhorne P, McKevittC, Bernhardt J, Rudd, A, Wolfe C. Anobservational study of acute stroke care infour countries: The European Registers ofStroke (EROS) Study. Cerebrovascular Dis28: 171-176. 2009Cumming T, Plummer P, Linden T andBernhardt J. Hemispatial neglect andrehabilitation in acute stroke. Archivesof Physical Medicine and Rehabilitation90:1931-1936.2009Sorbello D, Dewey H, Churilov L, ThriftAG, Donnan G, Bernhardt J. Very earlymobilisation and complications in the first3 months after stroke: Further results fromPhase II of A Very Early RehabilitationTrial (AVERT). Cerebrovascular Diseases28:378-383.2009Purvis T, Cadilhac D, Donnan G,Bernhardt J. Systematic review of processindicators: including early rehabilitationinterventions used to measure qualityof acute stroke care. International Journalof Stroke 4, 72-80.2009Bernhardt J, Thuy M, Collier J, Legg L.Very early versus delayed mobilisation.The Cochrane Library Issue 1. 2009Davis SM, Donnan GA. AntiplateletActivity. Stroke. [Editorial Material]. 2009Jun;40(6):2275-.Donnan G. Stroke: what can you do?Int J Stroke. [Editorial Material]. 2009Oct;4(5):313-.21

Clinical Trials andNeuroscience Trials AustraliaDonnan GA, Davis SM. IV and IAThrombolytic Stroke Strategies AreComplementary. Stroke. [EditorialMaterial]. 2009 Jul;40(7):2615-.Donnan GA, Davis SM. CombinedAspirin Plus Warfarin Recent Evidenceand Residual Questions. Stroke. [EditorialMaterial]. 2009 May;40(5):1946-.Donnan GA, Davis SM. The Ethicsof Thrombolytic Trials Beyond 3 (or4.5) Hours Randomized ControlledTrials Are Required to Change ClinicalPractice. Stroke. [Editorial Material]. 2009Apr;40(4):1545-.Davis SM, Donnan GA. Carotid-ArteryStenting in Stroke Prevention.N Engl J Med. [Editorial Material].2010 Jul;363(1):80-2.Davis SM, Donnan GA. Clot Retrieval forStroke ‘Yes We Can,’ but Priority for Trialsand Registries. Stroke. [Editorial Material].2010 Jan;41(1):196-.Purvis T, Cadilhac D, Donnan G,Bernhardt J. Systematic review of processindicators: including early rehabilitationinterventions used to measure quality ofacute stroke care. Int J Stroke. [Review].2009 Apr;4(2):72-80.Zhang WW, Cadilhac DA, Donnan GA,O’Callaghan C, Dewey HM. Hypertensionand TIA. Int J Stroke. [Review]. 2009Jun;4(3):206-14.22

Cognitive NeuroscienceOverview2009 was a year of development for theNeurocognitive Research Group in theFNI. We continued to strengthen linksbetween senior researchers and theirteams from the School of BehaviouralScience (University of Melbourne), theBrain Research Institute, the NationalStroke Research Institute, and theEpilepsy Research Centre. We increasedlinks between research projects and theclinical setting, with research at the BoxHill Hospital-based Eastern CognitiveDisorders Centre (ECDC) as well ascollaborations with researchers at AustinHealth, Royal Melbourne Hospital andthe FRONTIER group at the Prince ofWales Medical Research Institute. Weformed new collaborations with industry,contributing to a COGSTATE initiatedproject in which a community basedexamination of cognitive trajectory isstudied. The focus of our group remainsneuroscientific enquiry at a systemslevel in both basic and applied (clinical)fields, to facilitate the translation ofexperimental findings to improvedclinical practice using a sciencepractitionerapproach.Highlights include: well-attendedNeurocognitive Workshops arisingfrom monthly research meetings; ECDCsymposium at the Conference for ClinicalNeuropsychologists on the diagnosisand management of frontotemporaldementias; Churchill Fellowship for ECDCStaff member Dr Tracey Wardill, SeniorNeuropsychologist.Major Research ProjectsCortical remodelling associated withcarotid stenosisAmy Brodtmann, Fernando Calamante,Alan Connelly, Shawna Farquharson,David Abbott and Geoffrey DonnanCortical remodelling has beendemonstrated following stroke, often viafMRI studies. It is not fully understood ifsome of these changes are compensatoryor correlate with functional and cognitiveimpairments. There is evidence thathypoperfusion caused by vascular diseasedrives the recruitment of other areas ofthe brain not affected by arterial disease.We have studied a group of patientswith unilateral carotid arterial disease,and found that extensive remodellingis occurring in a region supplied by thenarrowed artery, the motor cortex. Thisstudy has provided new and importantinformation on the role of normal vascularperfusion on brain reorganisation, andfurther information on the effects ofhypoperfusion on cortical remodelling.Information on cortical perfusion iscritical, as it appears to be one of themost important determinants of corticalplasticity. In addition, understandinghow the brain responds to chronicischaemic stress may contribute to ourunderstanding of vascular cognitiveimpairment. These findings will now formthe basis of a larger longitudinal studyexamining cognition and brain volumeafter stroke.Is stroke degenerative? A longitudinalstudy of cognitive and changesregional brain volume following strokeAmy Brodtmann, Toby Cummingand Heath PardoeAlzheimer’s disease (AD) is the mostcommon form of dementia. While thecauses are not well understood, we knowfrom post-mortem studies that certainproteins (known as plaques and tangles)are found in the brains of patients withAD. These proteins act to acceleratecell death in the brain, which has theeffect of reducing brain volume overtime, a process known as atrophy. Strokeoccurs when blood supply to the brain isdisturbed, either by a blocked or a burstblood vessel. Historically, AD and strokehave been seen as separate entities.There may be reason to believe, however,that stroke has a role in the progressionof AD. Dementia caused by problemsin the brain’s blood vessels (vasculardementia) often co-exists with dementiacaused by AD. Research has shown thatAD patients with a history of stroke hadpoorer cognitive function than those whohad not suffered stroke. The questionremains as to whether stroke can triggerprogressive dementia and ongoing celldeath in the same way as AD. In thisproject, we propose a simple way toanswer this question. We will measurebrain volume (cortical thickness analysis)and cognitive function in stroke patientsseveral times over the first year afterstroke. These results will be comparedagainst healthy controls to determinewhether stroke is associated withprogressive reduction in brain volumeand cognitive decline.A neurobiological model of soundrecognition and processingNeil McLachlan and Sarah WilsonThe Object-Attribute model is nowsufficiently developed and detailedto enable specific hypotheses to betested in behavioural and neuroimagingparadigms. Predictions of thecomputational model have been testedagainst data from the literature. Newbehavioural tasks are currently beingdeveloped for EEG and fMRI studiesto validate specific neural mechanismsproposed in the model. In additionto understanding sound recognitionand identification, this project hasimplications for our understanding ofbehavioural habituation to auditoryenvironments, music processing, thedevelopment of new music technologies,sound recognition computer algorithms,and models of environmental noiseannoyance. It may also shed light onhallucinations experienced duringpsychosis and epilepsy.Neurobiological correlates ofexpressive and receptive musicalfunctionsSarah Wilson, David Abbott, Dean Lusher,Ellen Gentle and Graeme JacksonThis is a large-scale study investigatingthe relationship between musicand language using vocal singing(singing with lyrics) in both musicians,24

nonmusicians, and patients withtemporal lobe lesions. Vocal singingshares features common to music andlanguage, providing a unique opportunityto examine the relationships betweenhigher cortical functions. Currently it isnot clear, however, to what extent theyuse the same brain systems particularlyat the higher cortical level, and how thismight vary with singing ability. We haveused two neuroimaging paradigms toexamine the neural networks subservingsinging and language. We have foundthat the networks subserving singing andlanguage are proximally located in thebrain, and that their extent of corticaloverlap varies with singing expertise.Other Research ProjectsRepresentational neglect followingacute strokeTobias Loetscher, Amy Brodtmannand Mike NichollsSpatial neglect is a debilitating, butrelatively common, clinical disorderresulting from stroke. Neglect preventsthe patient from attending equally toboth sides of space – often resultingin bizarre symptoms such as ignoringpeople standing on the left anddressing just the right side of the body.Intriguingly, in addition to behave asthe left side of the world ceased toexist, these patients also ignore theleft side of imagined space. Whereasneglect research in the last decades hasmainly been focused on spatial deficitsin visuomotor space, in this project weexamine spatial impairments in imaginedspace. Using simple behavioural tasks,distortions in the representation ofimagined space is assessed in patientssuffering from neglect.Functional Neuroanatomy of DiscourseLeasha Lillywhite & Michael SalingBehavioural work on discourse productionsuggests a role for the temporal lobe inmessage-level representation. A studyof the impact of temporal lobe epilepsyon this network is in progress, as well asexamining patients with acquired aphasiasyndromes, particularly those withprimary progressive aphasia, part ofthe frontotemporal dementias.Alexia without agraphiaTobias Loetscher, Shawna Farqurson,Leasha Lillywhite and Amy BrodtmannThe incidence of Alzheimer’s pathologyon PET imaging in focal onset dementiaChristopher Rowe, Tracey Wardilland Amy BrodtmannFrontotemporal dementia registryAmy Brodtmann, Dennis Velakoulis,Richard Cotton, Colin Masters,Tiffany Cowie and David DarbyMirror neuron system function in theBroader Autism Phenotype: A windowinto the biology of empathySarah Wilson, Amee Baird, NeilMcLachlan, Alan Connelly andIngrid SchefferDeterminants of long-term outcomeafter acquired brain injury (ABI)Sarah Wilson, Joanne Wrench,Michael Saling and Graeme JacksonBiological & psychological predictorsof outcome after epilepsy surgerySarah Wilson, Michael Saling, JoanneWrench, Jeanette Lawrence, MarieO’Shea, Anne McIntosh and Sam BerkovicCharting the course of chronicity fromseizure diagnosisSarah Wilson, Michael Saling,Anne McIntosh, Sarah Velissaris,Mark Newton, Sam Berkovic, Gus Bakerand Anne JacobyUsing music to facilitate languagerecovery after strokeSarah Wilson, Dawn Merrett, IsabellePeretz and Graeme JacksonDeterminants of psychogenic seizuresSarah Wilson, Kim Gilson, Anne McIntosh,Ingrid Scheffer and Sam BerkovicPitch processing in musicians andnonmusiciansSarah Wilson, Neil McLachlan, DeanLusher, Dave Marco and Catherine MartinDevelopmental outcomes of childhoodonset temporal lobe epilepsySarah Wilson, Silvana Micallef,Asawari Killedar, Jacquie Wrennalland Simon HarveyCollaborationsEarly detection of Alzheimer’sdisease in a community setting usingcomputerised testing COGSTATEDavid Darby, Michael Woodward, LynetteMoore, Chris Rowe and Amy BrodtmannChanges in bone structure, leanmass, and glucose metabolismwithin the first six months post-stroke:a prospective studyKaren Borschmann, Marco Pang,Sandra Iliano-Burns, Elif Ekinci,Amy Brodtmann and Julie BernhardtProgressive aphasia and amyloiddeposition: a multidisciplinaryapproach to improving dementiadiagnosisJohn Hodges, Christopher Rowe,Olivier Piguet, Kerryn Pike, Kirrie Ballardand Victor VillemagneCollaboration on new projects withresearchers from the University ofMontreal, McGill University, and theMontreal NeurologicaI Institute hasculminated in the sabbatical placementof Associate Professor Wilson in the firstpart of 2010. Extensive collaborationsinterdigitating with Brain ResearchInstitute projects.25

Cognitive NeurosciencePublicationsBrodtmann, A Hashimoto’sencephalopathy and Down’s syndromeArch Neurol 2009 May;66(5):663-6Brodtmann, A, Puce, A, Darby, D, Donnan,G Regional fMRI brain activation doescorrelate with global brain volume BrainResearch 2009 Mar 9;1259:17-25Brodtmann, A, Puce, A, Darby, D, Donnan,G Post-stroke serial functional imagingreveals visual cortex reorganizationNeurorehab Neural Repair 2009 Feb23(2):150-159Cumming TB, Plummer-D’Amato P,Linden T, Bernhardt J (2009). Hemispatialneglect and rehabilitation in acutestroke. Archives of Physical Medicine andRehabilitation, 90, 1931-1936Joubert J, Reid C, Barton D, Cumming T,McLean A, Joubert L, Barlow J, Ames D,Davis S (2009). Integrated care improvesrisk-factor modification after stroke: Initialresults of the Integrated Care for theReduction of Secondary Stroke model.Journal of Neurology, Neurosurgery &Psychiatry, 80, 279-284.Random number generation inneglect patients reveals enhancedresponse stereotypy, but no neglect innumber space Loetscher T, Brugger P.Neuropsychologia. 2009 Jan;47(1):276-9.Neuropsychological and functional MRIstudies provide converging evidenceof anterior language dysfunction inBECTS. Lillywhite LM, Saling MM, HarveyAS, Abbott DF, Archer JS, Vears DF,Scheffer IE, Jackson GD. Epilepsia. 2009Oct;50(10):2276-84.Approachability and the amygdala:insights from Williams syndrome.Martens MA, Wilson SJ, Dudgeon P,Reutens DC. Neuropsychologia. 2009Oct;47(12):2446-53.Hippocampal volume and depression:insights from epilepsy surgery. WrenchJM, Wilson SJ, Bladin PF, Reutens DC.J Neurol Neurosurg Psychiatry. 2009May;80(5):539-44The neurocognitive components of pitchprocessing: insights from absolute pitch.Wilson SJ, Lusher D, Wan CY, DudgeonP, Reutens DC. Cereb Cortex. 2009Mar;19(3):724-32The experience of seizures after epilepsysurgery. Shirbin CA, McIntosh AM, WilsonSJ. Epilepsy Behav. 2009 Sep;16(1):82-5Personality development in the contextof intractable epilepsy. Wilson SJ, WrenchJM, McIntosh AM, Bladin PF, Berkovic SF.Arch Neurol. 2009 Jan;66(1):68-72.Cognitive complaints after a first seizurein adulthood: Influence of psychologicaladjustment. Velissaris SL, Wilson SJ,Newton MR, Berkovic SF, Saling MM.Epilepsia. 2009 May;50(5):1012-21.Characterising de novo depression afterepilepsy surgery. Wrench JM, Wilson SJ,O’Shea MF, Reutens DC. Epilepsy Res.2009 Jan;83(1):81-8Associate Staff and StudentsDr Amee Baird Postdoctoral FellowDr Toby Cumming Research FellowDr Leasha Lillywhite Postdoctoral FellowDr Tobias Loetscher Post-Doctoral ResearchFellow, University of MelbourneDr Dean Lusher Research FellowA/Prof Neil McLachlan Associate Professor(Acoustic and Auditory Modelling)Dr Ramesh Sahathevan Research Fellow,NSRIDr Joanne Wrench Postdoctoral Fellow26

Epilepsy— Epilepsy ImagingOverviewThere is a relatively high prevalence ofepilepsy in the general population (it hasbeen estimated that about 1 in every 140people suffer from epilepsy). Despite this,the mechanisms of seizure generationremain unclear. A significant proportionof people with continuing seizures areresistant to antiepileptic drugs; no viabletreatment options exist for some patients.Using combinations of neuroimagingtechniques at the FNI Austin precinct,we have made significant breakthroughsto help us understand the mechanismsof seizure generation. One of the keyshelping us unlock these secrets isfunctional magnetic resonance imaging(fMRI) and simultaneously acquiredelectroencephalography (EEG). Themethod that allows this technologyto be used simultaneously has beendeveloped at the BRI, and is the subjectof a current patent application. Structuraland physiological MRI methods are alsobeing developed and applied at BRI toinvestigate changes in brain networks thatoccur in epilepsy.Major Research ProjectsNHMRC Program Grant: “Epilepsy: abasic science to patient program abouthow gene changes cause seizures”Sam Berkovic, Graeme Jackson, JohnMulley, David Reutens, Steve Petrou,Ingrid Scheffer, Josef Getz, Alan Connellyand Anthony Waites.The epilepsy research Program consistsof the Epilepsy Research Centre (ERC) ofThe University of Melbourne (located atthe Austin precinct), the Epilepsy Imaginggroup and the MRI Development groupof FNI at the Austin precinct, the IonChannels and Human Disease groupat the Parkville precinct, and geneticsresearchers at the Women’s & Children’sHospital, Adelaide. It is arguably theworld leading program in epilepsygenetics and imaging, with major impactin understanding basic mechanisms andwith translational outcomes in patientmanagement. The collective researchrecord of the chief investigators showsvery high and increasing impact.We attribute this to the collaborativegain of our being part of a focussedand energetic team, bringing togetherfour nodes of independent highquality science.The achievements of this multi-disciplinaryteam include:• Revolutionary data leading to theepilepsies being regarded largely aschannelopathies.• Initial discovery and ongoingrefinement of imaging of hippocampalsclerosis by MRI.• Discovery of the role of the parietallobule in generalised epilepsy networks.• Solving the important challengeof “crossing fibres” in fibre-trackingMRI and applying the resultant novelmethods to clinical problems.• Providing the first evidence that, in amouse model of a human IGE, seizuresusceptibility is governed by bothdevelopmental and acute effects ofthe mutation.• Extensive validation of the GABRG2(R43Q) mutation discovered by us,studied by a vertically integratedbasic science program from cell toknock-in mice and correlated withhuman studies.Importantly, our genetic, imaging andbasic science discoveries are re-shapingthe clinical classification of epilepsies.Integration of the Research Teamsand ProgramOur greatest strength is the collaborativeand innovative working model oftranslational research that provides anintegrated approach to the mechanismsof disease. Many of our discoverieswould not have been possible withoutthe team’s collective abilities. We havelearned that it takes time for basicscientists to “get inside the head”of clinical researchers and vice versa.We have taken a disease-orientatedapproach to the important problemsof epilepsy and intellectual impairmentand have drawn together co-operative,collaborative and highly functionalmulti-disciplinary groups with proventrack records of productive science.Using multiple imaging modalities to“see” into the brainDavid Abbott, Richard Masterton,Kaushik Bhaganagarapu, Danny Flanagan,Radwa Badawy, Steve Fleming, JohnArcher, Simon Harvey, Alan Connellyand Graeme JacksonThe detection of changes in brain activityusing the electroencephalogram (EEG)has been a mainstay of the clinicaldiagnosis of epilepsy for decades. EEGinforms us about the existence and timingof abnormal synchronous neuronal events(including events that occur withoutany clinical symptoms); however EEG isunable to reveal the precise location ofthe brain activity. Magnetic ResonanceImaging (MRI) can also be used to imagebrain activity: functional MRI (fMRI) allowsthe exploration of the brain’s blood flowresponse to events, and is very goodat revealing the spatial location of theactivity. Compared to EEG however ithas rather poor time resolution. Marryingthe two techniques is technically verychallenging, for the MRI environmentconsists of very high static and changingmagnetic fields that usually swamp themicrovolt signals from the scalp EEG.We have developed a world-leadingmethod to overcome the technicallimitations allowing us to simultaneouslyacquire EEG and MRI. The device is nowin routine use at BRI, to investigate themechanisms of seizure generation(Figure 1). There is still much work tobe done to understand the informationthat we are now able to acquire, suchas determining how the location ofbrain activity associated with sub-clinicalabnormal electrical events in the brainmay relate to seizure generation itself.We hope that this work will eventually leadto a robust method for the localisation ofthe seizure focus in individuals with severeepilepsy, thus dramatically improvingthe chances of being able to surgicallyintervene to cure their condition.28

What is the sequence of events thatlead to a clinical seizure?Another promising approach is toinvestigate neurophysiological changesimmediately before the onset of a seizure.It is known that changes in brain activitycan occur minutes or even hours prior toclinical symptoms. We have successfullyused functional MRI to demonstrate areasof altered activity many minutes priorto seizure onset (Figure 2). Our resultsindicate that there are changes in thelead up to a seizure in areas that include,but are not limited to, the presumedseizure focus. The relationship of thesesignal changes to seizure generation, andhow this data might help to localise theepileptic network and “seizure focus” forsurgical treatment, is a complex questionthat we are actively pursuing.Brain networksRichard Masterton, Marnie Shaw,David Abbott, John Archer andGraeme JacksonFunctional magnetic resonance imaging(fMRI) was first demonstrated in humansin the early 1990’s, and has most oftenbeen used to identify focal regions ofthe brain associated with a particularcognitive function. However, functionalMRI can also be used to study extendednetworks of activity. Studies of so-called“functional connectivity” performed atBRI are helping map complex networksof interacting brain regions in healthypeople and in people with epilepsy. Weare interested both in technical aspectsof the method, as well as its potentialapplication in clinical research.Building global models of the brain’sfunctional networksRichard Masterton and Graeme JacksonAt a global level the brain is a singlehighly interconnected network thatintegrates separate functions that aredistributed across different specialisedbrain regions and sub-networks. Normalbrain function requires synchronisationand segregation of activities withindifferent parts of this network, anddisruptions in this balance are thoughtto be associated with many neurologicalconditions such as epilepsy.At the BRI we are using functionalconnectivity to probe the brain’sfunctional organization and buildcomplex models of its global networktopology based upon observedsynchronization and interaction betweenbrain regions. We are developingnew tools for creating and analyzingthese large network models. We haveconducted work that demonstratesthe brain’s network topology changesduring maturation from a centralized to adistributed organization. Continuing workis to investigate how different epilepsysyndromes – which often have specificage-dependent onset and remission –may interact with or be affected by thesebrain network changes.Figure 1Figure 1: An example of the abnormal electricalactivity in a patient’s brain detected by EEGrecorded from electrodes attached to thepatient’s scalp (the abnormal activity is circled).Simultaneously acquired functional MRI allows usto determine the location of blood flow changesassociated with the abnormal event. The locationis shown here in colour overlayed on image slicesof the brain in three different orientations.29

Epilepsy— Epilepsy ImagingFigure 2: Complex changes in fMRI signaltime-courses in the minutes prior to an epilepticseizure are revealed by several different analysismethods. Two time-courses (light blue and greenplots) of associated spatial maps extracted froman independent components analysis (ICA) of thefMRI time series are shown in the top-two plots.An hypothesis-driven analysis of significant changein mean signal in the minute prior to the seizurecompared to the minute commencing five minutesprior to the seizure reveals a hot-spot, shownin the bottom right image; mean time-coursesfrom a region of interest in the detected hot-spotcompared to a region on the other side of thebrain are shown in dark blue and pink respectively.Further exploration revealed a very focal regionadjacent to the hot-spot with a time-courseshowing abrupt signal intensity changes (orangeplot). All these time-course reveal changes atapparently synchronised time points: just over12 minutes prior to the seizure, andagain just over 6 minutes prior to the seizure,the ICA time-courses change frequency andamplitude, the very focal region shows abruptspiking and the mean signal in the “hot-spot”shows a marked divergence compared to theother side of the brain.Measuring changes in corticalexcitability in patients with epilepsyRadwa Badawy, Richard Macdonell,Samuel Berkovic and Graeme JacksonAt a biological level, epilepsy can beunderstood as a network of abnormallycommunicating brain areas, with a netincrease in brain cell activity. This theoryof increased excitability in epilepsy ismainly derived from animal studies andexperimental data and was until recentlyun-proven in humans because of the lackof the means to study this non-invasively.Transcranial magnetic stimulation (TMS)is a safe and non-invasive tool that canbe used to investigate the activity ofbrain cells and provide inferences on theexcitability of the brain in humans duringthe resting state (in between seizures).It is an excellent tool used to measurehuman brain cell excitability and thevarious circuits contributing to this. Mostof the previous TMS studies in epilepsyshowed conflicting results becausethey investigated patients with chronicepilepsy who were taking many antiepilepticdrugs. By studying patients withnew onset epilepsy who were not takingmedication, we found that there is analteration in brain cell activity associatedwith epilepsy with a net increase inbrain cell excitability. The pattern ofthis disturbance depended on the typeof epilepsy as it was on both sides ingeneralized epilepsies and confined toone side of the brain in focal epilepsies.We were also able to demonstrate thatthere was a cycle of change in excitability,as there was an additional increase inexcitability for 24 hours before a seizure,and a marked drop in excitability lastingfor 24 hours after a seizure before itreturned to baseline levels. Excitabilitywas also influenced by many factorsknown to provoke seizures. Our studiesshowed direct evidence of increasedexcitability early in the morning, premenstrualphase and sleep deprivation.When the patients started anti-epilepticmedication, this disturbance in excitabilitynormalized only if patients stoppedhaving seizures. Patients who continuedto have seizures showed progressivealterations in brain excitability. Thesefindings provide novel human insightsinto the mechanisms underlying epilepsy.Furthermore they are currently beingtranslated into clinical practice both in thediagnosis of new onset seizures and alsoto monitor responsiveness to treatment.Figure 230

Epilepsy— In Brain Markers of EpilepsyOverviewA phenotype is a physical characteristic,for example the colour of your eyes.Some phenotypes are associated witha disease, such as the short stature andflattened facial profile of those sufferingDown’s syndrome. An endophenotypeis in internal characteristic, such as theshape of your brain. At FNI, we using amultitude of neuroimaging sequencesincluding functional, structural andphysiological measures, to determinethe endophenotypes associated withparticular types of Epilepsy. These arethen combined with other data suchas genetic profiles, clinical symptomsand neuropsychological measures tohelp understand the disease. Particularfeatures associated with a condition maybe causative of seizures, a consequenceof seizures, neither or both. Researchersat FNI are helping determine thesignificance of identified features.Ultimately this will lead to improveddiagnosis and treatment of the disease.Figure 3Major Research ProjectsHow does disease affect brainfunction?Leasha Lillywhite, John Archer, MichaelSaling, Simon Harvey, David Abbott,Danya Vears (ERC), Ingrid Scheffer (ERC)and Graeme JacksonOne example of the endophenotypingstudies undertaken at BRI is thatof the specific epilepsy syndromeBenign Epilepsy with Centro-TemporalSpikes (BECTS). BECTS is a commoncause of epilepsy in primary schoolchildren. BECTS is characterised byfocal epileptic discharges (spikes)without frequent seizures or apparentbrain pathology. Children with BECTSalso experience cognitive difficultiesbut the relationship between thecognitive problems and the ‘spikes’was previously unknown. Participantsin our study completed a battery ofneuropsychological tests assessingintelligence, receptive and expressivelanguage, primary memory, new learning,academic attainment, and executivefunction. In addition, neuroimagingmeasures including structural, functional(including simultaneous EEG/fMRI) andphysiological (T2 relaxometry) wereacquired. Analysis of some of this datahas been completed and already wehave discovered that language-relatedactivation measured with fMRI is lesslateralized to the left hemisphere inanterior brain regions in patients relativeto the control group (Figure 3). Thisfinding is consistent with decreasedperformance in the BECTS groupcompared to the control group onthe neuropsychological measure mostdependent on the integrity of anterioraspects of the language axis, namely,sentence production. The result supportsthe view that BECTS is associated withlanguage difficulties that are regional,and anterior, in nature.Functional organisation of language inchildren with focal epilepsyRegula Everts*, Simon Harvey*, LeashaLillywhite, Jacquie Wrennall*, DavidAbbott, Linda Gonzalez*, Michael Kean*,Graeme D. Jackson and Vicki Anderson*(*Royal Children’s Hospital, Melbourne).We have used fMRI to map languageactivity in a group of children withepilepsy and a group of healthy children(some studied at FNI, others at the RoyalChildren’s Hospital in Melbourne). Wefound a correlation between languagelateralization (i.e. the side of the brainmost involved in language function)and verbal memory performance inpatients with left-sided epilepsy in thethree main areas of the brain responsiblefor language. Bilateral or right-sidedlanguage lateralization was associatedwith better verbal memory performance.Verbal memory performance madethe largest contribution to languagelateralization, whereas handedness andside of seizures did not contribute to thevariance in language lateralization. Thisfinding reflects the association betweenneocortical language and hippocampalmemory regions in patients with left-sidedepilepsy. Atypical language lateralizationis advantageous for verbal memoryFigure 3: Mean lateral indices (LIs) [ standarderror of the mean (SEM)] for the patient andcontrol group in each region of interest (toppanel). The regions of interest encompassed leftand right and middle frontal gyri (MFG), inferiorfrontal gyri (IFG), angular and supramarginal gyri(ANG), and posterior temporal lobe (Post. TEMP.)(Bottom panel).31

Epilepsy— In Brain Markers of Epilepsyperformance, presumably a result oftransfer of verbal memory function.In children with focal epilepsy, verbalmemory performance provides a betteridea of language lateralization thanhandedness and side of epilepsyand lesion.How does disease affect the structureof the brain?Heath Pardoe, Gaby Pell, David Abbottand Graeme JacksonStructural neuroimaging using magneticresonance imaging (MRI) is an importantdiagnostic and research tool, noninvasivelyproviding a three dimensionalview of the internal structure of the livinghuman brain. In some brain diseasesthere are obvious structural abnormalitiesthat can be seen using MRI. In many otherdiseases however we are discovering thatthere are subtle changes in brain structurethat cannot be seen by eye.Researchers at the brain research instituteare using MRI to investigate how brainstructures may be subtly altered in peoplewith conditions such as obstructive sleepapnoea and epilepsy, to assist in theunderstanding and better treatment ofthese conditions. For example, a deepbrainstructure often damaged in epilepsyis the hippocampus, and the volume ofthat structure in a patient compared tohealthy controls can be a phenotypicmarker of disease. We have beeninvestigating the efficacy of automatedcompared to manual segmentation ofthis structure, and have found that expertmanual hippocampal volumetry is moresensitive than the commonly advocatedautomated methods for the detectionof hippocampal atrophy associated withmesial temporal lobe epilepsy (Figure 4) –Heath to supply from his 2009 paper).The outcomes of our structuralimaging research will enable thescientific community to map patternsof structural change in brain tissue anddetermine the relationship betweenthese structural changes and alteredfunction of pathological tissue. Forexample, “Long-term outcomes inchildhood-onset epilepsy” is an NIHfunded project currently underway. Thisis an ongoing prospective cohort studyof 613 children recruited when firstdiagnosed with epilepsy. The projectaddresses two issues: 1) More than halfof adults with intractable epilepsy whoare treated surgically have epilepsyof childhood onset, yet, prospectivestudies find few children with surgicallytreatable intractable epilepsy. 2) Inotherwise neurologically normal adultswith childhood onset epilepsy, many ofwhom experience complete remissionof seizures and come off medication,educational and social outcomes areimpaired relative to non-epilepsy controls.The role of BRI in the project is to applyadvanced image analysis techniques tostructural MRI scans acquired from a largesubset of the original cohort.These methods provide insight intolinks between brain structure and social,educational and health related outcomesin patients with childhood-onset epilepsy.We have applied these techniquesto localize structural brain changes inchildhood absence epilepsy, and addressmethodological issues involved in theanalysis of imaging data acquired atmultiple imaging centres.Is Obstructive Sleep Apnea AssociatedWith Changes In Brain Morphology?Heath Pardoe, David Abbott, Gaby Pell,Ferghal O’Donoghue (Austin Health),Melinda Jackson (Austin Health), MaryMorrell (Imperial College, UK), DougCorfield (Keele University, UK) andGraeme JacksonObstructive sleep apnea is a disordercharacterized by obstructed breathingduring sleep. The disorder causesintermittent hypoxia and frequentarousals from sleep, which can havea considerable negative impact inlifestyle. Moderate sleep apnea has beenestimated to affect up to 24 percentof middle-aged men. There is a wellrecognisedlink between obstructivesleep apnea and cognitive dysfunction.This project was a collaborative studybetween FNI, the Department ofRespiratory and Sleep Medicine atthe Austin Hospital and the Sleep andVentilation Unit at the Royal BromptonHospital in the UK. We used an imageanalysis method known as voxel-basedmorphometry to investigate changes inbrain structure associated with obstructivesleep apnea. The voxel-based approachwas modified using an approachpioneered by the Brain Research Instituteto allow for comparison of MRI-dataacquired from multiple imaging centres.The results of the study suggested thatthe structure of the cerebellum maybe affected in people with obstructivesleep apnea.Figure 432

Neuroanatomical changesin childhood-onset epilepsyHeath Pardoe, Gaby Pell, David Abbott,Anne Berg (Northern Illinois University)and Graeme JacksonThe prognosis for childhood-onsetepilepsy is complex. Many patients withchildhood-onset epilepsy stop havingseizures for an extended period, only tohave seizure activity re-occur several yearslater, which requires surgical treatment.These cases are rarely imaged early in thecourse of their epilepsy. Little is knownabout these epilepsy cases which may beassisted by early intervention to preventthe re-occurrence of seizures.In this study, we are using advancedimage analysis techniques to identifyneuroanatomical changes in over 300people with childhood-onset epilepsy.The use of quantitative analysis ofMRI-based neuroimaging data will helpto determine how changes in brainstructure related to epilepsy affect social,educational and health-related outcomesin these patients.Image analysis methods utilised in thisstudy include:(i) Hippocampal Volumetry(ii) Voxel-based morphometry(iii) T2-relaxometry(iv) Cortical thickness analysisThe involvement of FNI in such a uniquelarge-scale study of childhood epilepsyprovides valuable insight into themechanisms by which epilepsy can affectthe quality of life of people burdenedwith this disease.Figure 5Quantitative imaging of corticalabnormalities in extratemporal epilepsyHeath Pardoe and Graeme JacksonEpilepsy is often associated withabnormalities in cortical structure. Theidentification of abnormal cortex isimportant in the pre-surgical assessmentof intractable epilepsy patients becausethe chances of seizure freedom followingsurgical resection of the suspectedepileptogenic region are higher if a lesionhas been identified on neuroimaging. Inthis project we developed a method forthe identification of cortical abnormalitiesin patients with extratemporal epilepsyusing cortical thickness analysis ofstructural MRI, alignment of the structuralMRI scans using high-dimensional nonrigidregistration, and statistical analysis toobjectively identify cortical abnormalitiesin individuals. The method developedin this project was successfully used toobjectively identify cortical abnormalitiessuch as focal cortical dysplasias andgliomas in epilepsy patients.Can we diagnose disease by imagingbrain physiology?MRI scanners are extremely flexibleimaging machines, capable of noninvasivelydetecting tissue characteristicsand even determining the chemicalmakeup of internal organs in the body.Current research at FNI includes thedevelopment of improved imagingsequences and automated analyticalmethods to detect subtle changes inbrain tissue characteristics in patientswith various diseases.T2-relaxation time: An indicator of thehealth of brain tissueThe measurement of T2 relaxation time(“T2 relaxometry”), has been establishedas a reliable tool for the quantitativemeasure of tissue abnormalities intemporal lobe epilepsy. At BRI themeasurement of the T2 is achieved usingan optimized pulse sequence that hasbeen developed in our Institute. We havealso developed a novel analysis methodthat we call voxel based relaxometry (VBR)which provides an automated objectivedescription of areas of T2 abnormalityin groups of patients with epilepsycompared to control patients. However,most current clinical neuroimaging isundertaken using qualitative rather thanquantitative modalities. At BRI we havedeveloped and validated a method forthe analysis of inherently qualitativeimages. Unlike quantitative modalitieswhere a consistent quantitative measureof a physical property of the tissue isobtained (eg T2-relaxometry measuresthe tissue’s T2-relaxation time), qualitativeimages are designed to produce relativecontrast between tissue types or activity,but the intensity values can be far fromconsistent between imaging sessions(eg T2-weighted MRI results in visiblecontrast between grey and whitematter in the brain, however thetissue’s T2-relaxation time cannot bedetermined from these images). Ouranalysis method for these images iscalled Voxel Based Iterative SensitivityAnalysis (VBIS). It optimally adjustsglobal signal of qualitative images tominimise global signal variation betweenimages, maximising sensitivity to thelocal (regional) signal differences of realinterest. We have validated the methodfor T2-weighted images, as shown infigure 6.Figure 5: VBIS-T2 is a method for the quantitativeanalysis of qualitative T2-weighted images. Herewe show a comparison of the method with thegold-standard fully-quantitative technique voxelbasedrelaxometry (VBR) in a group of 24 patientswith left hippocampal sclerosis, compared to 97healthy controls. The mean of all T2-weightedimages is shown in greyscale. Yellow overlayindicates T2 change detected by both VBR andVBIS-T2. Green indicates change detected onlyby VBIS-T2. Red indicates change detected onlyby VBR. VBIS-T2 was able to correctly identify thedominant area of T2 signal difference in the lefttemporal lobe.33

Epilepsy— In Brain Markers of EpilepsyPublicationsAbbott DF, Pell GS, Pardoe H, JacksonGD. Voxel-Based Iterative Sensitivity(VBIS) analysis: Methods and a validationof intensity scaling for T2-weightedimaging of hippocampal sclerosis.Neuroimage 2009;44(3):812-819. 2009Badawy R, Macdonell R, JacksonGD, Berkovic SF. Why do Seizures inGeneralized Epilepsy often occur in theMorning? Neurology 2009, 73(3):218-22.2009Badawy R, Macdonell R, Jackson GD,Berkovic SF. The peri-ictal state: corticalexcitability changes within 24H of aseizure. Brain 2009; 132:1013-1021. 2009Calamante F, Connelly A. PerfusionPrecision in Bolus-Tracking MRI:Estimation Using the Wild-BootstrapMethod. Magnetic Resonance inMedicine 2009; 61:696-704. 2009Calamante F, Connelly A, van Osch MJP.Nonlinear Delta R-2* Effects in PerfusionQuantification Using Bolus-Tracking MRI.Magnetic Resonance in Medicine 2009;61:486-492. 2009Close TG, Tournier JD, Calamante F,Johnston LA, Mareels I, Connelly A. Asoftware tool to generate simulated whitematter structures for the assessment offibre-tracking algorithms. Neuroimage2009; 47(4):1288-1300. 2009Datta S, Hart GK, Opdam H, GutteridgeG, Archer J. Post-hypoxic myoclonicstatus: the prognosis is not alwayshopeless Crit Care Resusc (2009) 11(1)39-41. 2009De Tiege X, Connelly A, Liegeois F,Harkness W, Clark CA, Chong WK,Gadian DG, Cross JH. Influence of motorfunctional magnetic resonance imagingon the surgical management of childrenand adolescents with symptomatic focalepilepsy. Neurosurgery 2009; 64(5):856-64.2009Flanagan D, Abbott DF, Jackson GD.How wrong can we be? The effect ofinaccurate mark-up of EEG/fMRI studiesin epilepsy. Clinical Neurophysiology2009, 120(9):1637-47. 2009Gardner H, Lawn N, Fatovich DM, ArcherJS. Acute hippocampal sclerosis followingecstasy ingestion Neurology (2009) 73(7)567-9. 2009Gilmore R, Sakzewski L, Boyd R.Systematic review and meta-analysis oftherapeutic management of upper-limbdysfunction in children with congenitalhemiplegia. Pediatrics 2009; 123:1111-1122. 2009Lillywhite LM, Saling MM, Harvey AS,Abbott DF, Archer JS, Vears D, SchefferIE, Jackson GD. Neuropsychological andfunctional MRI studies provide convergingevidence of anterior language dysfunctionin BECTS. Epilepsia 2009, 50(10):2276-2284. 2009O’Reilly M, Bollmer B, Vargha-KhademF, Neville BGR, Connelly A, Wyatt J,Timms C, de Haan M. Ophthalmological,cognitive, electrophysiological andMRI assessment of visual processingin preterm children without majorneuromotor impairment. DevelopmentalScience 2009; 1-14. 2009Pardoe H, Pell G, Abbott D, JacksonGD. Hippocampal volume assessmentin temporal lobe epilepsy: how good isautomated segmentation Epilepsia 2009,50(12):2586-92. 2009Pillay N, Berkovic, SF, Fitt G, Fabinyi G,Myles T, Jackson GD. Parahippocampalepilepsy with subtle dysplasia: a causeof ‘imaging negative’ partial epilepsyEpilepsia 2009, 50(12):2611-8. 200934

Epilepsy— Ion Channels and Human DiseaseResearch OverviewEpilepsy is a debilitating brain diseasethat afflicts 10 percent of the populationat some stage. Three percent of theworld’s population is currently diagnosedwith epilepsy. Anti-epileptic drugs havesignificant shortcomings and almost athird of patients are unable to achieveadequate seizure control. At present,surgery is the main recourse for thesepatients, who often live for 10 or moreyears with untreatable epilepsy prior tosurgical intervention. New therapeuticsare urgently needed to not only treatpatients that don’t respond to currentanti-epileptic drugs but to also reduce theserious side effects of these medications.Our laboratory has taken the approachthat novel opportunities for therapeuticintervention will arise by; 1) the creationof syndrome-specific epilepsy modelsbased on human genetic lesions and;2) a detailed analysis of the fundamentalmechanisms that underlie disease genesisand progression in these models.To achieve these goals we employ amultidisciplinary approach that combinesmolecular biology, biophysics, computermodelling, single cell and brain sliceelectrophysiology, macro and microhistological digital imaging, EEG, unitrecording and in vivo patch clamp inbrains and behavioural analysis of mice.By studying the effects of epilepsy genemutations at several levels of functionalorganisation, we can validate our modelsagainst the human conditions and thendelve into the mechanisms of seizuregenesis. We have strong commercial linksso that discoveries in the lab can morereadily benefit patients.Major Research ProjectsDevelopment of syndrome specificmodels of human epilepsySyndrome specific models of epilepsyare the key to understanding thefundamental origins of this disease andto the development of better therapeutictreatments. The goal of this project isto develop genetically modified mousemodels of specific human epilepsysyndromes. These studies begin withthe function validation of epilepsycausing gene mutations found in patientpopulations with a collaborative team ledby Prof Samuel Berkovic. The majorityof these mutations are found in a classof proteins called ion channels. Thesemembrane spanning proteins creategated switches in neurons, controlexcitability of neurons and underliethe basic functioning of the brain. Weassess the effects of mutations by usingpatch clamp electrophysiology, twoelectrode voltage clamp and a range ofcell biological methods. Once diseasecausing mutations are identified in humanstudies, mouse models are createdthat harbour the identical mutations.Validation of these models requiresdetailed study at the molecular, neuronal,network and whole animal level. Wesearch for similarities in cellular behaviourand seizure activity between our miceand the patients with the mutation asthis provides us with greater confidencethat fundamental mechanisms of seizuregenesis may be shared. Our searchfor the mechanism of epileptogenesisrequires the use of miniaturised singleneuron recording devices for use inunrestrained and awake animals as wellas use of in vivo patch clamp recordingsin anesthetised mice. We couple thesestudies with EEG and behavioural testsas our markers of syndrome type. Wehave used molecular biological methodsto engineer switching of gene mutationson and off in living mice to investigatethe study of direct versus developmentaleffects of mutations. We envisage thata range of different models will have tobe created to cover the major refractoryepilepsies seen in man. One major activityis the phenotypic characterisation ofeach of these three models. With thesemodels in hand our studies should revealmechanisms of seizure genesis, thusproviding fertile ground for developmentof novel therapeutics.Neuron specific inducible modelsof human familial epilepsyChris Reid and Steven PetrouIn this project, we created and validateda novel mouse model in which we cantemporally control the expression of ahuman epilepsy mutation. To do this,we used a gene silencing Neomycinresistance cassette which normallysuppresses the expression of themutation, however, with the use of adrug called doxycycline, we can switchon the mutation by in vivo excision ofthe silencing Neomycin cassette. Themutation is in the GABA receptor genewhich has major impact during braindevelopment as well as playing animportant direct inhibitory role in theadult brain. By expressing the mutationafter brain development has completed,we have addressed a central issue ofepilepsy genetics which is whetherepilepsy gene mutations cause epilepsyby perturbing brain development orthey directly impact brain functionto result in seizures. So far, we havepreliminary evidence that the mutationhas developmental effects which worsenseizures later in life. More work is currentlyunder progress to study the possibledevelopmental effects of this mutation.Other uses of this model includeexpressing the mutation in differentcell types in the CNS to determine theneuronal components that drive seizurephenotypes. Mouse lines that express inexcitatory neurons (EMX1) and a separateline that express only in inhibitoryneurons (DLX) are currently being breedwith our conditional model and testingof seizure susceptibility is underway. Withthe creation of this novel model, we nowhave the ability to gain precise controlover temporal and spatial aspects ofepilepsy gene expression.Dissecting the mechanisms of clinicalheterogeneity in familial epilepsyVerena Wimmer, Chris Reid,and Steven PetrouThe genetic basis of epilepsy is knownto be highly complex. It displays geneticheterogeneity where different genes cancause the same epilepsy phenotype aswell as clinical heterogeneity where thesame gene can cause different epilepsyphenotypes. In the GABA receptorgene, two mutations have been foundthat result in a different phenotype.One mutation is seen in patients withchildhood absence epilepsy (CAE) andfebrile seizures, which are relatively mildforms of epilepsy, and in one family amutation in the same gene results insevere myoclonic epilepsy of infancy(SMEI) which is an extremely severecondition with a poor prognosis. We areinterested in the clinical heterogeneityof the GABA receptor gene and wish tounderstand how two different mutationsin the same gene can cause two verydifferent phenotypes. We have madea mouse model with the mutation that35

Epilepsy— Ion Channels and Human Diseasegives rise to the less severe forms ofepilepsy and we are now making asecond mouse model with the GABAreceptor mutation that gives rise toSMEI. Future work on these mice modelswill help us identify and understand thepathways leading to different types ofepilepsy. Identification of the point ofphysiological divergence that resultsin these two conditions may yieldimportant clues as to the fundamentalmechanism of epileptogenesis in thesedifferent syndromes.Sodium channels splicing and seizuresusceptibilityElena Gazina and Steven PetrouClinical data have identified connectionof a L1563V mutation in the SCN2Aprotein (a sodium channel protein) tochildhood epilepsy. The SCN2A proteinhas 2 isoforms, neonatal and adult. Ourdata has shown that when expressed inHEK293 cells, the neonatal channel isless excitable than the adult form, andthe L1563V mutation of the mutant formresults in an increase in excitability toadult protein levels. Our hypothesisis, therefore, that alternative splicingmay be critical in reducing excitabilityin the infantile brain and susceptibilityto seizures. We are currently geneticallyengineering mutant mice with permanentneonatal or adult SCN2A genes totest this hypothesis and clarify thedevelopmental roles of these isoforms.Hyperpolarisation activatedcation current in familial andacquired epilepsyChristopher Reid, Alison Marie Philips,Verena Wimmer and Steven PetrouThere is an obvious clinical need forimproved therapeutics that will only berealised by a better understanding ofepilepsy. A key goal in our understandingof this disease is to define the molecularparticipants involved in the epileptogenicprocess and attribute functionalconsequences to them. One candidateclass of protein is the hyperpolarisationactivated cation channels (HCNs). Thereare a number of studies that have showedchanges in HCN expression and functionduring the development of epilepsy, buttheir impact on seizure susceptibilityremains unclear. This motivates ourhypothesis: that selective up and downregulation of HCN expression canmodulate seizure susceptibility. The aimof this project is to take advantage ofstate-of-the-art recombinant viral genedelivery methods to change HCN1 andHCN2 levels. Injection of these tools intodifferent parts of the brain will allow usto determine if changing HCN proteinexpression can produce or ‘rescue’epilepsy. We have now validated theHCN shRNA tool in vitro and in vivostudies are looking at how reducingthe HCN1 channel changes neuronalnetwork properties. Also, together withour collaborators we are the first tofind a mutation in the HCN gene thatmay act to increase the susceptibilityof an individual’s likelihood of gettingepilepsy. Using a medium-throughputelectrophysiological assay we havedemonstrated that this variant changeschannel function. We are using thisvalidated assay to determine if otherHCN channel variants change function.The research aim is to determine theimpact of this mutation on channelfunction and how this may changeseizure thresholds.Interneurons in epilepsy – are theythe real culprits?Verena Wimmer, Kay Richards, ChristopherReid, Elisa Hill and Steven PetrouHarmonious functioning of the brainhinges upon accurate excitatory andinhibitory actions from pyramidalneurons and GABAergic interneurons,respectively. Seizures arise fromhyperexcitability of the brain, which maymanifest from inadequate GABAergicinhibitory actions. Little is known aboutinhibitory interneurons and epilepsy,but it is commonly thought thatseizures arise from ”disinhibition” ofthe neuronal network. It is now clearthat this concept is oversimplified.Recent evidence demonstrates thatin epileptic brain tissue, GABAergicnetworks undergo complex rewiring atanatomical, physiological and functionallevels. These novel results indicate thatthe role of GABAergic interneuronsin epilepsy is more complicated thancan be explained just by a reduction orabsence of inhibitory activity. The aim ofthis project is to investigate interneuronproperties in an epileptic mousemodel using immunohistochemicaland electrophysiological techniques.Our laboratory has produced, andis characterising, the first transgenic(knockin) mouse model carrying a humanepilepsy mutation (R43Q) on the GABA Areceptor γ2 subunit (GABRG2(R43Q)).GABAergic interneurons make uparound 20% of neurons in the brain andare difficult to identify. To overcomethis limitation, in this project, we willcross our existing epileptic mouse witha transgenic mouse that expressesgreen fluorescent protein (GFP) inall interneurons, thus allowing theidentification of interneurons in the brain.Using the R43Q mouse model of absenceepilepsy, this project will investigateinterneuron properties in three regionsinvolved in the thalamocortical circuit,the reticular thalamic nucleus (NRT)and the somatosensory cortex. Theneurochemical properties, electricalproperties and connectivity of thesegreen fluorescing interneurons will becharacterised in wild-type and epilepticmice to determine whether interneurondevelopment (migration, morphologyand syaptic bouton density) and functionis altered in our epilepsy mutant mice.Analysis of GABA-releasing inhibitoryinterneurons in R43Q mice will provideinsights into the underlying mechanismsleading to the generation of absenceseizures. This study will provide arationale as to how the R43Q mutationin the GABA A receptor γ2 subunit alterselectrical network activity in the brain tocause epilepsy.Febrile Seizures and temperaturedependentGABA A receptor traffickingdeficiencies in the R43Q mutantElisa Hill, Susan Hosie and Steven PetrouFebrile seizures affect as many as onein 15 children. Epilepsy has long beenthought to be related to alterationsin GABAergic transmission in thethalamocortical circuitry of the brain.Our laboratory has generated a knockinmutant mouse with an arginine replacedby a glutamine residue (R43Q) in thegamma2 subunit of the GABA A receptor.This mouse model exhibits a phenotypesimilar to that seen in a large Australianfamily demonstrating a high incidenceof febrile seizures carrying the samegene mutation. The gamma2 GABA Areceptor subunit is critical for normalreceptor trafficking. Furthermore, febrileseizures have been correlated with atemperature dependent reduction of cellsurfaceexpression of GABA A receptors36

containing impaired gamma2 subunitfunction. This project aims to assessthe temperature dependence of R43QGABA A receptor trafficking in corticalslices by examining GABA A receptormediatedsynaptic events using the patchclamp electrophysiological technique.It is hypothesised that at elevatedtemperatures, GABA A receptor-mediatedsignal detection will be reduced,resulting in increased excitability in thethalamocortical circuitry. In the presenceof glutamatergic receptor antagonists,synaptic GABA A currents will be recordedfrom cortical pyramidal cells in preheatedor room temperature acute slices frommutant and wild type animals. In addition,the expression of the gamma2 subunitin preheated versus control corticalslices will be assessed by quantitativeimage analysis of immunocytochemicaldistribution patterns of cell-surfaceexpressed receptor. The identificationand analysis of a temperature-sensitivereduction in receptor expression in acutecortical slices may lead to importantadvances in drug targets and possibletherapies in the field of febrile epilepsy.Environmental influences of seizuresusceptibilityChris Reid and Steven PetrouAbsence epilepsies are highly heritable,but it is well known that environmentaleffects such as decreased vigilanceand voluntary hyperventilation mayinduce absence seizures. However, theusual physiological or environmentalprecipitants of absence seizures are notfully understood. Recently, there hasbeen renewed interest in the relationshipof glucose concentration to absenceseizures with the discovery that absencesseizures, and other generalized seizuretypes, occur in patients with impairedtransport of glucose across the bloodbrain barrier due to mutations in SLC2A1that encodes GLUT1, the major brainglucose transporter. These seizuresoccur in the classical severe GLUT1encephalopathy, but also in milder casesthat have otherwise typical idiopathicgeneralized epilepsy associated withSLC2A1 mutations. Interestingly, patientswith GLUT1 deficiency respond to oralglucose with significant improvement inboth clinical seizures and EEG findings.Blood glucose levels fluctuate naturallyand whether reductions in levels influenceabsence seizure threshold is unclear. Thisquestion is difficult to study in childrenwith the disease. We are addressing thisissue experimentally in genetic micemodels with varying susceptibility toabsence seizures. Low blood glucose canreduce seizure threshold in geneticallypredisposed animal models suggestingthat they be considered as a potentialenvironmental risk factor in absenceepilepsy patients. This is exciting workthat may have immediate impact onclinical practice.Genetic background influences onthalamocortical nuclei in a mousemodel of absence epilepsyChristopher Reid and Steven PetrouRecently, a mutation was found tobe linked to a generalised epilepsysyndrome with Childhood AbsenceEpilepsy (CAE) in a large Victorianfamily. Absence seizures are defined asseizures that result in sudden onset ofbrief periods of unconsciousness. Theoccurrence of these seizures commonlydecline after six years of age. Interestingly,a mouse model incorporating thismutation developed by our grouprecapitulated this epileptic condition,confirming the causal role of this mutationin CAE. Previously, we have demonstratedthat the mutation results in impairedGABA A receptor trafficking, which resultsin a deficit in cortical inhibition. Ourlab now focuses on relevant neuronalnetworks and in vivo in order to relatethese molecular deficits with the clinicalpathology of the disease. In familiesharbouring the GABA A receptor γ2(R43Q) mutation, the presence of themutation is linked to Childhood AbsenceEpilepsy. However, there is a strongclinical heterogeneity and not all subjectswith the mutation exhibit an epilepticphenotype. In order to investigate theinfluence of genetic background on thefinal phenotype of this mutation, themutation was bred into two differentgenetic backgrounds; the C57/B6 andthe DBA/2J mice strains. The geneticbackground of the animal harbouring themutation also exerted a strong influenceon the epileptic phenotype. Mutant C57/B6 mice only exhibited 4-10 seizures perhour with each seizure lasting only 0.5-1s.In contrast, mutant mice in the DBA/2Jbackground exhibited 40-150 seizuresper hour with each seizure lasting 1-3s.To investigate the physiological differencebetween these two strains responsiblefor contrasting seizure susceptibility, atechnique called Patch Clamp recordingwas used to record synaptic signals inbrain slices to characterise how differentcells communicate with each other. Wefocused our efforts on the cortex andthe thalamus, two large brain regionsknown to play an important role inAbsence epilepsy. Interestingly, theDBA strain showed a large increasein synaptic inhibition in the thalamusbut not the cortex. Inhibition in thethalamus is essential in driving thisregion of the brain into synchronizedepileptic activity. This inherent differencebetween the mice strains may thereforeresponsible for the different seizureseverity they develop as a result of themutation. This suggests that although asingle mutation may be a primary causefor the development of epilepsy, othergenetic factors strongly influence thefinal clinical outcome. We are takinga genetic approach to determiningwhat differences exist between theC57 and DBA stains that define seizuresusceptibility. DBA by C57 crosses thathabour the γ2 (R43Q) mutation havevaried seizure susceptibility potentiallyallowing us to map to genes of interest.An extension of this approach is currentlybeing completed by our collaboratorProf Wayne Frankel (Jacksons Laboratory,USA). Here the R43Q mutation isbeing breed into a range of differentmice species and seizure susceptibilitymeasured. Both approaches hope todiscover new seizure modulating genesthat will give some insight into the basisof clinical heterogeneity.Connecting the dots from molecularlesion to network seizure in epilepsyEvan Thomas and Steven PetrouRecent advances in the genetics ofepilepsy have lead to the discoveryof many mutations that cause seizuresyndromes. Almost without exceptionthese mutations are in ion channels –molecules that reside in the cellmembrane and conduct current that isthe basis of electrical signalling in thenervous system. In our laboratory we usea variety of techniques to understand howthese molecules are different from thosein humans without epilepsy. One of theseis to artificially express these ion channelsin special cell lines and measure thedifferences in their electrical propertiescompared to ion channels from healthy37

Epilepsy— Ion Channels and Human Diseasepeople. These differences are small andsubtle and it is often far from clear howthe cause the brain to become proneto seizure.One way to make the casual connectionbetween molecular deficit and changesin brain dynamics is through the useof computer models. Data collectedfrom cell line studies can convertedto mathematical models of the ionchannel in question. These modelscan then be incorporated into modelsof single neurons and used to predictdifferences in sensitivity of these neuronsto interactions from other neurons.Then these the neuron models canbe incorporated in network modelswith thousands of neurons to try tounderstand what triggers neurons.Understanding why some brains are moreliable to seizure than others will enableus to develop not only better treatmentsfor seizure prevention but will help usunderstand why brains become epilepticin the first place.New methods for exploringcomplex modelsJordan Chamber, Steven Petrou andEvan ThomasMany diseases, including epilepsy, arecaused by subtle biophysical changesin the response of proteins. Thus, inorder for computational models to bepredictive they must capture sufficientdetail that these subtle changes areevident. This poses a new problem in thatthese models then have large numbers ofparameters, many of which are not wellestablished or not mutually consistent.New methods are needed to explorelarge computationally demandingmodels. To develop these methods, weare using a well respected thalamocorticalmodel already exists. We have taken theNEURON (an open source simulationpackage) port of this model and updatedthe model to included significant newfindings from the last 5 years. Briefly, themodel consists of 13 neurons types fromthe cortex and thalamus, with realisticmorphology and connectivity. The neuronmodels express a realistic complementof voltage and calcium sensitive ionchannels, AMPA, NMDA, GABA Amediated synaptic conductances andgap junctions. While this is not acomplete model of the cortex andthalamus, it contains the importantcomponents believed to play a significantrole in the generation of epilepticseizures. Indeed, the original model andour updated model (from preliminaryresults) are able to display both normaloscillations and epileptic oscillations.The goal of this project is to usetechniques to intelligently explore theparameter space of the model and linkthis to data from experiments in animalmodels. The model contains severalthousand parameters describing neuronmorphology, probabilities of connectionsbetween different neuron types,biophysical properties of ion channels,distribution of ion channels and dynamicsof postsynaptic responses. Some ofthese parameters are perturbed by thedisease. Experimental studies indicatethat epileptic oscillations are caused bydeficits in inhibitory transmission in thecortex and changes in the distributionof inhibitory interneurons in the cortexand thus provide important clues as towhich subset of parameters is important.Exactly which parameters are perturbedby the disease is unknown, but thesedata reduce the number of potentiallyimportant parameters to around 40. Thisparameter space will be explored usingthe Nimrod toolkit (http://messagelab.monash.edu.au/Nimrod) for workflowmanagement of complex parametricexperiments. This is a unique tool forworking with parametrically complexmodels and we believe this is the firstapplication of these techniques toquestions in neuroscience. We will usethe Nimrod/E tools to create a fractionalfactorial design for the parameter spacesearch. This works as follows. If N valuesof each parameter are to be tested thena brute force approach will require N 40runs to test every parameter combination.A factorial design is based on the ideaof first testing single parameter changes,followed by two simultaneous parameterchanges, then three and so on. Typicallyhigher order interactions have negligibleeffect. In this way, the number of runscan be reduced to around 20,000 toexamine 1, 2 and 3-way parameterinteractions. Exploring a complete set of4-way interactions is not computationallyfeasible, however data from this projectwill be used to determine if 4-wayinteractions are likely to be important andto specify a subset of interactions thatmay be worth pursuing.Multi-modal imaging analysis of mousemodels of human epilepsy syndromesKay Richards, Gary Egan, David Reutensand Steven PetrouHow does a gene mutation, representingdysfunction at the molecular level,lead to altered whole brain functionsuch as epilepsy?To begin to tackle this question we areutilising mouse models of inheritedepilepsy syndromes, combined withmulti-modal imaging. Our researchgoal is to identify macroscopic andmicroscopic structural changes in thebrain which may result in the generationof seizures. Our methodology consistsof three parts: 1) MRI; 2) Histology and 3)High resolution 3D brain reconstruction.Part 1 includes analysis of grossanatomical structure of whole mousebrain using magnetic resonance imaging(MRI). Evidence from human MRI studiesof patients who carry the ion channelmutation linked to generalised epilepsywith febrile seizures plus (GEFS+), showdecreased brain volume in the corpuscallosum. Our preliminary results fromT2 weighted images of mice which carrythe same gene mutation identified inGEFS+ suggest altered brain volumecompared to control animals. Parts 2and 3 of the project involve collection ofhistological stained serial sections of thesame brain used to obtain MR imaging.The 30µm thick sections, imaged atx4 magnification have been used toproduce high resolution histologicaldata. All animals will have co-registrationof MRI and histological sections as a 3Dvolume data set (collaboration with ProfDavid Reutens). The purpose of this istwofold: first, co-registration of MRI and3D volume data set provide important invivo information about changes in brainvolume in the epilepsy model. Second:comparisons of MRI and histology ofthe brain identify structural alterationsthat are a consequence of histologicalprocessing. In addition; mapping highresolution information from sub structuresin the brain back into a 3D space willidentify specific regions which are alteredin gene mutant animals, for examplechanges in brain nuclei and white mattertracts. Overall, this multi-modal imagingapproach will provide valuable anatomicalstructural information and lead the wayfor future analysis.38

Altered temperature sensitivityof familial sodium channel mutationsin epilepsy as a unifying mechanismfor genesis of febrile seizuresEvan Thomas and Steven PetrouGeneralised Epilepsy with FebrileSeizures Plus (GEFS+) type 1 ischaracterised by a C121W point mutationin the ß1 subunit of the voltage gatedsodium channel. People afflicted withthis condition display seizures initiatedby fever up until six years of age, andthen display febrile or afebrile seizuresinto adulthood. Previous publishedstudies using cell lines have shownthat this sodium channel mutationalters the functionality of the channel.The aim of this project is to study thesodium channel’s kinetics propertieswith changes in temperature thatcorrespond to both normal and feverbody temperatures in humans. Data sofar show changes in channel kinetics aswell as differing temperature sensitivitiesof the mutated channel compared tothe normal channel. Preliminary data atfever temperature indicate that changesin temperature sensitivity may cause ahigher susceptibility to seizures. Futurework will entail action potential firingproperty studies of neurons in C121Wmouse models as well as investigating anew technique called Dynamic Clampingthat may enable action potential firingproperties of mutated ion channels to beunderstood in simple cultured cells suchas human embryonic kidney (HEK293)cells instead of neurons.Kinetic models of ion channel gatingfor gene environmental interactionsEvan Thomas and Steven PetrouOne growing area of interest ispharmacoresistance where variationin the patients genome prevents thesymptoms from being controlled withdrugs. Some pharmacoresistance willbe due to variation in ion channelresponses to drugs, indeed this maybe the same variation that causedthe disease. Other environmentalinteractions, such as temperature andph, are triggers of seizures in individualswith heightened sensitivity. In order, toincorporate these effects into predictivesingle neuron and network model, highquality mathematical models of the ionchannels response to both endogenousand exogenous stimuli is needed. This issurprisingly difficult to do properly, evenin the case of Hodgkin-Huxley models.The reasons are that gradient followingalgorithms suffer from two problems.Firstly, they are easily fooled by localminima which is a point in parameterspace that only minimizes the error ina small region. Secondly, they handlediscontinuities and singularities poorly,both of which are present in exponentialbased rate functions. To overcomethese difficulties, a strategy needs tobe developed to generate a good firstguess of the solution. The strategy usedis to initially fit a Hodgkin-Huxley model,convert that to its equivalent kineticmodel, remove the relationship betweenrate functions and finally use a patternsearch algorithm to perform the final fit.High content high throughput analysisof gene variation in epilepsy.Evan Thomas and Steven PetrouThe aim of this proposal is to understandthe functional consequences of geneticvariation found in sodium channel alphasubunits in order to develop a deeperunderstanding of genetic risk in familialepilepsy. The recent explosion of variantdata coming from patients with epilepsyhas identified sodium channels, and inparticular SCN1A, as key determinant ofsusceptibility. Studies of rare monogenicepilepsies have provided unequivocalevidence of how single gene variantscan lead to seizures but have not shedmuch light on the genetic architectureof the more common forms of epilepsy.However, little is know of how geneticvariation seen in the population alterschannel function and impact risk.Developing a full picture of “risk” requiresan assessment of the pathogenicity ofvariants and for ion channels this meanselectrophysiological analysis. To datemost studies have focussed on analysingone or at most several variants, typicallyfrom a monogenic case. Two technicalissues arise in the planning of large scaleanalysis of sodium channel variants. Oneis that risk alleles will likely have a smallerfunctional deficit than monogenic diseasealleles, and the second is that traditionalpatch clamp studies are too slow andexpensive to be able to deal with thenumbers of variants needed to establisha complete picture of risk for a givenallele. Fortunately, recent developmentsin the high content high throughputanalysis of ion channels have providedsolutions to both these problems. Solvingthe first problem requires large numbersof replicates and reduce scatter of thedata in order to achieve statistical powerto call small changes in ion channelproperties, the current generation ofion channel screening devices can dothis. Establishing a more completepicture of risk requires an understandingof the impact of genetic variation atmultiple levels of brain organisation,from molecular deficit to impact onbrain networks. To achieve this we willincorporate the changes in sodiumchannel function into models of neuronsand networks and assess the impact ofvariants on these higher level functions.Using this combined approach wehope to develop a more thorough andpredictive understanding of the geneticrisk for epilepsy.Ion transport blockers as antiviralsElena Gazina and Steven PetrouWe have demonstrated that a number ofamiloride derivatives have antiviral activityin tissue culture against viruses belongingto two medically important genera ofthe Picornavirus family: coxsackievirus B3(CVB3, causes myocarditis) and humanrhinovirus 2 (HRV2, a common cold virus).The compounds were more effectiveagainst CVB3 than against HRV2. Ourdata suggested that the antiviral activityagainst coxsackievirus B3 was due toinhibition of viral RNA replication, withthe antiviral target likely to be a viralprotein. In the case of HRV2, virus releasefrom infected cells was also inhibited.We have generated amiloride-resistantCVB3 mutants by passaging the virusin the presence of the compound. Weare currently sequencing the genomesof the resistant virus isolates to identifymutation(s) causing resistance, andthus identify viral protein(s) targetedby amiloride and its derivatives. Oncethe protein is identified, the studieswill be conducted to elucidate thedetailed mechanism of antiviral activity.We have analysed anti-CVB3 activityand cytotoxicity of 29 compoundsbelonging to a large group of chemicals,acylguanidines, which includes amiloridederivatives. The resulting structure-activitydata were used to delineate medicinalchemistry strategy aimed at improvingthe antiviral activity and cytoxicity ofthe compounds.39

Epilepsy— Ion Channels and Human DiseaseNeurosteroids as potential analgesics;what is the mechanism of action?Brett A. Cromer and Steven PetrouPain, particularly chronic pain, can bedebilitating and is a major health issuethroughout the world. Opioids are themost effective analgesics available andare widely used to alleviate severe painbut are not effective against all formsof pain. They also suffer from unwantedside-effects including nausea, vomiting,respiratory depression and dependence.Opioids act largely by decreasing theexcitation of dorsal horn neurons thattransmit pain signals. GABA receptorsregulate this transmission, suggestingthat GABA modulators may providealternative analgesics. Indeed manyGABA modulators do relieve pain but areunusable for systemic treatment of painbecause they also cause depression ofbrain activity and anaethesia. Recently, aneurosteroid has been found to relievepain by acting on GABA receptors,without causing anaesthesia, offeringpromise for the development of newanalgesics. To capitalise on this promisewe first need to understand which specificGABA receptor is targeted, as thereare many types, and the precise natureof the active compound, as metabolicprocessing is required for activity. Wehave recently instigated a researchproject to answer these questionsusing a combination of metabolictracing, electrophysiological testing ofcompounds on a panel of recombinantlyexpressed GABA A receptors andphysiological testing in geneticallymodified mice. We have found that,relative to alphaxaone, alphadolone hasGABA A receptor subunit selectivity thatis consistent with recent evidence for thesubunits that are important for spinal painpathways. These findings may explain theanalgesic effects of alphadolone.Modulation of ClC-1 by ATP;a role in fatigue?Brett A. Cromer, Steven Petrou andGraham LambClC-1 has a similar role in muscle tothat of the GABA A receptor in thebrain, mediating inhibitory currentsthat modulate muscle excitability andactivation. Mutations in the ClC-1 genecan lead to hyper-excitability and stiff orrigid muscles in the disease, myotoniacongenita. We have shown recently thatregulatory domains of ClC-1 controlchannel function in response to ATP (theenergy currency of the cell) and acidosis,a mechanism that may be important formuscle fatigue during intense exercise.We are investigating further the molecularmechanism underlying this modulationand, in collaboration with Graham Lambat Latrobe University, its physiologicalsignificance.The Role of P2X7 Receptors inNeuroinflammation and DegenerationBen Gu, Steven Petrou and James WileyThe P2X7 receptor is a ligand-gatedcation channel highly expressed onmonocytes, macrophages and microgliawhich mediates the pro-inflammatoryeffects of extracellular ATP. Prolongedexposure of these cells to ligand leadsto channel dilatation and massive K+efflux which in turn is a co-stimulus forthe secretion of cytokines, interleukin-1and interleukin-18 which perpetuateinflammation. Whole animal evidencesuggests that P2X receptors on microglialcells in the brain and spinal cord playa major role in amplifying the damagedue to inflammatory or traumatic lesions.Several polymorphic variants of theP2RX7 gene are associated with muchenhanced permeability responses of thereceptor and these polymorphic variantshave been associated with greaterinflammation in an autoimmune disease,Sjogrens Syndrome. Other reports haveassociated bipolar disorder with a gainof function polymorphism in the P2RX7gene which is consistent with the currentconcept of bipolar disorder as an ionchannelopathy . While there is a clearrole for P2X7 in inflammation, emergingevidence suggests a second unsuspectedfunction for P2X7 in innate immunity.Thus macrophages or other cell typestransfected with P2X7 can recognize andengulf foreign particles, gram positiveand negative bacteria and even apoptoticautologous lymphocytes. This phagocyticfunction of P2X7 is only observed inthe absence of ATP and allows for theremoval of foreign material or apoptoticdebris in the absence of inflammation. This unique property of the P2X7receptors may be attributed to an arrayof anti-parallel beta-pleated sheets inthe extracellular domain providing anextensive interacting surface with theparticle to be engulfed. However thesignificance for the nervous systemof these varying roles for P2X7 remainsto be elucidated.Staff ListProfessorial FellowsJames S. Wiley MDPost Doctoral FellowsAlison Clarke BSc (Hons) PhD(University of Masschusetts)Elena Gazina BSc PhD (Moscow) Dip Ed (Mon)Elisa L. Hill AMusA BSc (Hons) PhD (Melb)Susan Hosie PhDAlison Marie Phillips PhD (Melb)Christopher Reid BPharm (Tas) BSc(Hons)PhD (ANU)Kay L. Richards BAppSci (Hons) (RMIT)PhD (Melb)Evan A. Thomas BSc (Hons) (Melb) MA (SUNYat Stony Brook) MAppSci(IT) (RMIT) PhD (Melb)Verena C. Wimmer DiplBiol (University ofRegensburg) PhD (University of Heidelberg)Brett A. Cromer BSc (Hons) PhD (ANU)Han Shen Tae PhDRobert Richardson BSc (Hons) PhD (Melb)Jordan Chambers BSc (Hons) PhD (Melb)Ernesto Vargas BSc PhD (Melb)Research AssistantsChantal Trager BSc (Melb)Emily Cunningham BSc (Hons) MelbNathan Myhill BASebastian Jungnickel PhDNerida Taylor BScLynley Cordeiro BScStudentsTae Hwan KimBryan Tangwen LeawMohd Zulqisti Mohd ZulkifliByron ScafEva SoMegan OlivaRosemary Harty40

David KaplanRobert HatchMelody Yuen Sin LiOral PresentationsSteven PetrouAmerican Epilepsy Society AnnualMeeting, Boston, USA, 2009American Epilepsy Society AnnualMeeting, USA, 2009Californian Institute of Technology,Pasadena, USA, 2009Centre for Neuroscience, Australia, 2009Curtin Conference, Australia, 2009Epilepsy Program Retreat, 2009Department of Genetics,University of Melbourne, Australia, 2009Department of Physiology,University of Melbourne, Australia, 2009Eccles Institute of Neuroscience Seminars,Australia, 2009Jasper’s Basic Mechanisms of theEpilepsies Workshops, Yosemite,USA, 2009NeuroParkville 2009, Australia, 2009Repligen Inc, Massachusetts, USA, 2009Sepracor Inc, Massachusetts, 2009The Jackson Laboratory,Maine United States, 20096th Joint Annual Meeting of the German,Austrian and Swiss Leagues of theInternational League, Germany, 2009Brett CromerInitiated meeting and Chaired OrganisingCommittee for Curtin Conference on IonChannels and Transporters, Canberra,April 2009MyoNaK, International muscle sodiumand potassium meeting, Palm Cove,Qld. Aug. 2009, “CBS-domain mediatedmodulation of skeletal muscle ClC-1chloride channels by ATP and acidosis –a role in fatigue?”Epilepsy Program Retreat, HepburnSprings Aug 2009 “Structure andFunction: the GABA A receptor and earlyonset epilepsies GEFS + – associatedγ2R43Q mutation”.Melbourne Neuroscience SymposiumNov. 2009 “Structure and Function:the GABA A receptor and early onsetepilepsies GEFS + – associated γ2R43Qmutation”.Chair “Ion Channels and Transporters”session at combined AuPS/ANS meetingsFeb 2010Evan ThomasIon channel variants in the dentate gyrus.Epilepsy program mini-retreat 2009Heat opens sodium channels. Epilepsyprogram retreat 2009The biophysical basis of epilepsy.Swinburne University of Technology 2009Modelling risk in inherited epilepsy.University of New South Wales 2010Verena WimmerHuman epilepsy mutation revealsfunction of Scn1b sodium channel subunitin the axon initial segment. AnnualRetreat of the Epilepsy Research Centre,2009, DaylesfordQuantitative morphology of molecularand neuronal changes in IGE‘IGE day’ of the Epilepsy Research Centre,2009, MelbourneCellular mechanism of hyperexcitabilityin the SCN1B (C121W) mouse model ofhuman epilepsy.Australian Neuroscience Society AnnualMeeting, 2009, CanberraChris ReidApril 2009: Department of Physiology,Monash UniversityMolecular mechanisms underlyingepilepsyMay 2009: Department of Anatomy andcell biology, University of MelbourneGenetics of epilepsy; Humanised modelsEpilepsy program retreat 2009HCN: a genetic cause of epilepsy?November 2009 NeuroscienceSymposium:A new model of Dravet SyndromeDecember 2009 Dept of Physiology,Penn State University, USABenzodiazepine pharmacology in ahumanised mouse model of epilepsyMajor Collaborative LinksNationalProf Sam BerkovicAustin HospitalAnalysis of gene mutations infamilial epilepsyProf John MulleyWomen’s and Children’s Hospital,AdelaideValidation of epilepsy causing mutationsProf David ReutensMonash Medical CentreConstruction of a 3D probability mapof the mouse brainProf Graeme JacksonFlorey Neuroscience InstitutesValidation of a novel NMR imagingmodalityA/Prof David AndersonBurnett InstituteDevelopment of anti-picornoviralsProf Joe LynchMolecular Pharmacology of GlycineReceptorsProf Lloyd HollenbergUniversity of MelbourneQuantum decoherence methods foroptical imaging of neuronal activityInternationalA/Prof Matt JonesUniversity of Madison, Wisconsin, USABiophysical analysis of GABAA receptorfunction in brain slices in the R43Q mousemodel of generalised epilepsyProf Kai KailaUniversity of Helsinki, FinlandpH dependence of axon initialsegment functionProf Tallie BaramUniversity of California, Irvine, USAAnalysis of Ih mutations in epilepsy41

Epilepsy— Ion Channels and Human DiseaseProf Holger LercheUniversity of Ulm, GermanyFunctional analysis of genetic variationin IGEProf Wayne FrankelJackson Laboratories, Bar Harbour, USAGenetic and functional analysis ofepilepsy in the mouse modelProf Heinz BeckUniversity of Bonn Medical Centre,GermanyAxon initial segment dysfunctionin epilepsyPatentsSteven Petrou and Evan Thomas.Modulation of an ion channel or receptor.2008906148. AUSTRALIA. 2009. PCTPublicationsLiao Y, Anttonen AK, Liukkonen E, GailyE, Maljevic S, Schubert S, Bellan-Koch A,Petrou S, Ahonen VE, Lerche H, LehesjokiAE. SCN2A mutation associated withneonatal epilepsy, late-onset episodicataxia, myoclonus and pain. Neurology,In Press.New therapeutic opportunities inepilepsy: a genetic perspective. Reid CA,Jackson GD, Berkovic SF, Petrou S. Petrou.Pharmacology and Therapeutics. In Press.Liao Y, Deprez L, Maljevic S, Pitsch J,Claes L, Hristova D, Jordanova A, Ala-Mello S, Bellan-Koch A, Blazevic A,Schubert A, Thomas EA, Petrou S, BeckerAJ, De Jonghe P, Lerche H. Molecularcorrelates of age-dependent seizures inan inherited neonatal-infantile epilepsy.Brain (2010) 133:1403-14Dibbens LM, Reid CA, HodgsonB, Thomas EA, Phillips AM, Gazina E,Cromer BA, Clarke AL, Baram TZ, SchefferIE, Berkovic SF, Petrou S. Augmentedcurrents of an HCN2 variant in patientswith febrile seizure syndromes. Annals ofNeurology, (2010) 67:542-6Thomas EA, Reid CA, Petrou S. Predictionby modelling that epilepsy may becaused by very small functional changesin ion channels. Arch Neurol (2009)66:1225-32Bajor A, Ung K-A, Öhman L, SimrenM, Thomas EA, Bornstein JC, SjövallH. Indirect evidence for increasedmechanosensitivity of jejunalsecretomotor neurons in patients withidiopathic bile acid malabsorption.Acta Physiol, (2009) 197:129-37.Thomas EA, Reid CA, Petrou S. Mossyfibre sprouting interacts with sodiumchannel mutations to increase dentategyrus excitability. Epilepsia, (2009)51:136-45Thomas EA, Hawkins RJ, Richards KL,Xu R, Gazina EV, Petrou S. Heat opensaxon initial segment sodium channels:A febrile seizure mechanism? Annals ofNeurology, (2009) 66:219-226.Wimmer VC, Reid CA, Mitchell S,Richards KL, Scaf BB, Leaw BT, Hill EL,Royeck M, Horstmann MT, Cromer BA,Davies PJ, Xu R, Lerche H, Berkovic SF,Beck H, Petrou S. Axon initial segmentdysfunction in a mouse model of geneticepilepsy with febrile seizures plus. J ClinInvest. 2010 Jul 12. pii: 42219.Wimmer VC, Reid CA, So EY, BerkovicSF, Petrou S. Axon initial segmentdysfunction in epilepsy. J Physiol. 2010Jun 1; 588(Pt 11):1829-40.Nicolazzo JA, Steuten JA, CharmanSA, Taylor N, Davies PJ, Petrou S. Brainuptake of diazepam and phenytoinin a genetic animal model of absenceepilepsy. Clin Exp Pharmacol Physiol.2010 May; 37(5-6):647-9.Gazina EV, Richards KL, MokhtarMB, Thomas EA, Reid CA, Petrou S.Differential expression of exon 5 splicevariants of sodium channel alpha subunitmRNAs in the developing mouse brain.Neuroscience. 2010 Mar 10;166(1):195-200.Goldschen-Ohm MP, Wagner DA, PetrouS, Jones MV. An epilepsy-related regionin the GABA(A) receptor mediateslong-distance effects on GABA andbenzodiazepine binding sites. MolPharmacol. 2010 Jan; 77(1):35-45.Bird MK, Reid CA, Chen F, Tan HO, PetrouS, Lawrence AJ. Cocaine-mediatedsynaptic potentiation is absent in VTAneurons from mGlu5-deficient mice. Int JNeuropsychopharmacol. 2010 (2):133-41.Dibbens LM, Harkin LA, Richards M,Hodgson BL, Clarke AL, Petrou S, SchefferIE, Berkovic SF, Mulley JC. The role ofneuronal GABA(A) receptor subunitmutations in idiopathic generalizedepilepsies. Neurosci Lett. 2009 Apr 10;453(3):162-5.Reid CA, Berkovic SF, Petrou S.Mechanisms of human inheritedepilepsies. Prog Neurobiol. 2009 Jan 12;87(1):41-57.Zheng T, Clarke AL, Morris MJ, Reid CA,Petrou S, O’Brien TJ. Oxcarbazepine, notits active metabolite, potentiates GABAAactivation and aggravates absenceseizures. Epilepsia. 2009 Jan; 50(1):83-7.Barrett GL, Reid CA, Tsafoulis C, Zhu W,Williams DA, Paolini AG, Trieu J, MurphyM. Enhanced spatial memory andhippocampal long-term potentiation inp75 neurotrophin receptor knockout mice.Hippocampus. 2010 Jan; 20(1):145-52.Adams BE, Reid CA, Myers D, Ng C,Powell K, Phillips AM, Zheng T, O’BrienTJ, Williams DA. Excitotoxic-mediatedtranscriptional decreases in HCN2channel function increase networkexcitability in CA1. Exp Neurol. 2009 Sep;219(1):249-57.Monif M, Reid CA, Powell KL, Smart ML,Williams DA. The P2X7 receptor drivesmicroglial activation and proliferation: atrophic role for P2X7R pore. J Neurosci.2009 Mar 25; 29(12):3781-91.Powell KL, Cain SM, Ng C, SirdesaiS, David LS, Kyi M, Garcia E, Tyson JR,Reid CA, Bahlo M, Foote SJ, SnutchTP, O’Brien TJ. A Cav3.2 T-type calciumchannel point mutation has splicevariant-specificeffects on function andsegregates with seizure expression in apolygenic rat model of absence epilepsy.J Neurosci. 2009 Jan 14; 29(2):371-80.Meyer HS, Wimmer VC, Hemberger M,Bruno RM, de Kock CP, Frick A, SakmannB, Helmstaedter M. Cell Type-SpecificThalamic Innervation in a Column of RatVibrissal Cortex. Cereb Cortex. 2010 Jun24. [Epub ahead of print].Meyer HS, Wimmer VC, Oberlaender M,de Kock CP, Sakmann B, HelmstaedterM. Number and Laminar Distribution ofNeurons in a Thalamocortical ProjectionColumn of Rat Vibrissal Cortex. CerebCortex. 2010 Jun 24. [Epub ahead of print].42

Wimmer VC, Bruno RM, de Kock CP,Kuner T, Sakmann B. Dimensions of aProjection Column and Architecture ofVPM and POm Axons in Rat VibrissalCortex. Cereb Cortex. 2010Wimmer VC, Möller A. High-resolutionconfocal imaging in tissue. Methods MolBiol. 2010; 611:183-91.Chen, X., Cromer, B., Webb, T. I., Yang, Z.,Hantke, J., Harvey, R. J., Parker, M. W. &Lynch, J. W. Dihydropyridine inhibition ofthe glycine receptor: Subunit selectivityand a molecular determinant of inhibition.Neuropharmacology. 2009: 56, 318-327.Chen, X., Cromer, B. A. & Lynch, J. W.Molecular determinants of beta-carbolineinhibition of the glycine receptor. JNeurochem 2009: 110, 1685-94.43

Genomic Disorders Research CentreOverviewThe Genomic Disorders Research Centre(GDRC) was formed to lead the world ingenetic research focussing on mutationand its effects on human well-being. TheGDRC was the first and remains the onlyCentre world wide to focus on mutations.In this post Human Genome Project erathe Genomic Disorders Research Centrestrives for excellence, focusing on genemutation, its cause, documentation,collection and consequences. The Centrehas numerous national and internationalactivities such as courses, workshopsand a high impact factor and high profilegenetics journal Human Mutation.HistoryThe Centre was established in January1996 to undertake medical and scientificresearch in the area of genetic mutationdetection and its impact upon the humancondition. Originally it was administeredunder the auspices of the St. Vincent’sInstitute of Medical Research and calledthe Mutation Research Centre.In November 1997 it changed its nameto the Foundation for the Detectionof Genetic Disorders, although from ascientific point of view it was still referredto as the Mutation Research Centre. InJanuary 1998 it became independentfrom the Institute and during 2000 anew name was adopted, with the intentof more readily identifying the Centre’sbroadening research effort. Since thistime the Centre has been known as theGenomic Disorders Research Centre. In2001 it adopted a neurological focus andwas a partner in the successful NationalNeuroscience Facility grant; this helpedforge closer liaisons in 2002 with theNeurosciences Victoria/Australia. During2003, the administrative office movedto the National Neuroscience Facilityin Carlton and the Genomic DisordersResearch Centre became a fully ownedsubsidiary company of the Howard FloreyInstitute and presently the GDRC is aGroup of the Howard Florey Institute.The GDRC is currently focusing all itsresources on the Human Variome Project.PurposeThe GDRC continues to make inroads onone of the central international projectsdeveloped by the GDRC, to establishthe visionary Human Variome Project inMelbourne and the high ranking journalHuman Mutation.The Human Variome ProjectThe Human Variome Project is the globalcommunity effort to collect, curateand make accessible information onall genetic variations affecting humanhealth. It was initiated at a meeting inMelbourne, Australia in 2006 by a groupof internationally prominent researchersand clinicians working with geneticvariation in order to formalise existinglinks and provide a central coordinatingbody. Since then, the concept of theHuman Variome Project has evolved andmatured to become a partnership ofcountries and organisations working tocreate the systems necessary to ensurethat all genetic variations of clinicalrelevance are collected, curated andmade freely available.The GDRC hosts the InternationalCoordinating Office of the HumanVariome Project. This work enables globalstandards systems and collaborators tobe developed for gene variation datacollection specifically those causinginherited disease from all genes in allcountries. The GDRC has encouragedmany initiatives to be established inparticular that of the HVP NeurogeneticsConsortium which is working towards thecollection of genetic data implicated inmany neurological disorders.Australian Node – Human VariomeProjectOver the past year the GDRC has beengranted funding to develop the softwareand systems for the HVP Australian Node.This work will enable gene variationdata to be collected from AustralianLaboratories and this information willallow enhanced diagnosis abilitiesfor Australian clinicians when treatingpatients with many inherited cancersand debilitating disorders. This workis intended to form a model for theestablishment of data collections inother countries.Mutation Databases, Human GenomeVariation Society and Human MutationThe experience of initiating the journalHuman Mutation, led to the realizationthat a huge amount of data will beuncovered regarding inherited disease.This led to the set up of the HumanGenome Variation Society (HGVS). TheHuman Variome Project grew out of this(see above).Tim Smith has embarked on a PhDproject to examine the role of thedatabase curator in the construction,maintenance and operation of geneticvariation databases. These databasesprovide vital information to clinicians anddiagnostic specialists on the frequency,clinical effect and genetic consequencesof numerous variations in our geneticmakeup and are frequently being usedas clinical tools for the management andtreatment of patients. However, despitetheir importance and frequent use, datamanagement and preservation strategieshave never been fully explored in thisparticular field. This project is attemptingto define a standard methodology for thecuration of these vital resourcesOther activitiesThe GDRC administers the secretariatfor the HGVS and its two internationalmeetings per year and the Editorial officefor Human Mutation.Information About TheAchievements/ AchievementsIn The 2009 Calendar YearWe have been organising the thirdHuman Variome Project meeting heldunder UNESCO Patronage 10-14 May2010 at the UNESCO Headquarters, Parisand the Australasian Mutation Detectionmeeting to be held in Tasmania, August,2010, among others.We received funding for the developmentof software and systems to establish theAustralian Node of the Human VariomeProject from the Federal GovernmentNeAT grant scheme.The Human Variome Project office hasassisted the establishment a local HVPgroup in Jeddah.44

International meeting organised by theGDRC in 2009“Towards Establishing Standards”, HumanVariome Project Forum, Vienna, Austria,May 22.“Genome Wide Association Studies: thelast mile problem”, Human GenomeVariation Society Scientific Meeting,Vienna, Austria, May 23.Mutation Detection 2009, X InternationalSymposium on Mutation in the HumanGenome, Paphos, Cyprus, May 28-June 1.“Spotlight on Neurogenetics”, HumanVariome Project Forum, Honolulu, USA,October 19.Human Genome Variation Society BoardMeeting, Honolulu, USA, October 19.“Impact of Next Generation Sequencing”,Human Genome Variation SocietyScientific and Annual General Meeting,Honolulu, USA, October 20.Notable PublicationGENETICS. The Human Variome Project.Richard G. H. Cotton, Arleen D. Auerbach,Myles Axton, Carol Barash, Samuel F.Berkovic, Anthony J. Brookes, John Burn,Garry Cutting, Johan T. den Dunnen,Paul Flicek, Nelson Freimer, Marc S.Greenblatt, Heather J. Howard, MichaelKatz, Finlay A. Macrae, Donna Maglott,Gabriela Möslein, Sue Povey, RajkumarS. Ramesar, Carolyn S. Richards, DanielaSeminara, Timothy D. Smith, María-JesúsSobrido, Jan Helge Solbakk, Rudolph E.Tanzi, Sean V. Tavtigian, Graham R. Taylor,Joji Utsunomiya, Michael Watson. Science,2008. 322(5903): p. 861-2Publications in 2009Somatic mutation databases as tools formolecular epidemiology and molecularpathology of cancer: proposed guidelinesfor improving data collection, distribution,and integration. Olivier, M., Petitjean, A.,Teague, J., Forbes, S., Dunnick, J. K., denDunnen, J. T., Langerod, A., Wilkinson,J. M., Vihinen, M., Cotton, R. G. andHainaut, P. Human Mutation, 30(3):275-282, 2009.Sharing data between LSDBs and centralrepositories. Johan T. den Dunnen, RolfH. Sijmons, Paal S. Andersen, MaunoVihinen, Jacques S. Beckmann, SandroRossetti, C. Conover Talbot Jr., Ross C.Hardison, Sue Povey, Richard G.H. Cottonand the Human Genome Variation Society(HGVS). Human Mutation, 30(4):493-495,2009.Planning the Human Variome Project:The Spain report. Kaput, J., Cotton, R.G., Hardman, L., Watson, M., Al Aqeel,A. I., Al-Aama, J. Y., Al-Mulla, F., Alonso,S., Aretz, S., Auerbach, A. D., Bapat,B., Bernstein, I. T., Bhak, J., Bleoo, S.L., Blocker, H., Brenner, S. E., Burn, J.,Bustamante, M., Calzone, R., Cambon-Thomsen, A., Cargill, M., Carrera, P.,Cavedon, L., Cho, Y. S., Chung, Y. J.,Claustres, M., Cutting, G., Dalgleish, R.,den Dunnen, J. T., Diaz, C., Dobrowolski,S., Dos Santos, M. R., Ekong, R., Flanagan,S. B., Flicek, P., Furukawa, Y., Genuardi, M.,Ghang, H., Golubenko, M. V., Greenblatt,M. S., Hamosh, A., Hancock, J. M.,Hardison, R., Harrison, T. M., Hoffmann,R., Horaitis, R., Howard, H. J., Barash, C. I.,Izagirre, N., Jung, J., Kojima, T., Laradi, S.,Lee, Y. S., Lee, J. Y., Gil-da-Silva-Lopes, V.L., Macrae, F. A., Maglott, D., Marafie, M.J., Marsh, S. G., Matsubara, Y., Messiaen,L. M., Moslein, G., Netea, M. G., Norton,M. L., Oefner, P. J., Oetting, W. S., O’Leary,J. C., de Ramirez, A. M., Paalman, M. H.,Parboosingh, J., Patrinos, G. P., Perozzi,G., Phillips, I. R., Povey, S., Prasad, S., Qi,M., Quin, D. J., Ramesar, R. S., Richards,C. S., Savige, J., Scheible, D. G., Scott, R.J., Seminara, D., Shephard, E. A., Sijmons,R. H., Smith, T. D., Sobrido, M. J., Tanaka,T., Tavtigian, S. V., Taylor, G. R., Teague, J.,Topel, T., Ullman-Cullere, M., Utsunomiya,J., van Kranen, H. J., Vihinen, M., Webb,E., Weber, T. K., Yeager, M., Yeom, Y.I., Yim, S. H. and Yoo, H. S. on behalfof contributors to the Human VariomeProject Planning Meeting HumanMutation, 30(4):496-510, 2009.A novel and simple method of screeningcompounds for interaction with DNA:A validation study. Adel Garas, ElizabethWebb, Vino Pillay, Donald MacPhee,William Denny, Hilary Zeller andRichard Cotton. Mutation Research,Genetic Toxicology and EnvironmentalMutagenesis, 678(1):20-9, 2009.Capturing all disease-causing mutationsfor clinical and research use: Toward aneffortless system for the Human VariomeProject. Richard G.H. Cotton, Aida I. AlAqeel, Fahd Al-Mulla, Paola Carrera,Mireille Claustres, Rosemary Ekong,Valentine J. Hyland, Finlay A. Macrae,Makia J. Marafie, Mark H. Paalman,George P. Patrinos, Ming Qi, RajkumarS. Ramesar, Rodney J. Scott, Rolf H.Sijmons, María-Jesús Sobrido, MaunoVihinen and members of the HumanVariome Project Data Collection fromClinics, Data Collection from Laboratoriesand Publication, Credit and IncentivesWorking Groups. Commentary, Geneticsin Medicine, 11(12):843-9, 2009.The Chemical Cleavage of Mismatchfor the Detection of Mutations in LongDNA Fragments. Tania Tabone, GeorginaSallmann, and Richard G. Cotton. SingleNucleotide Polymorphisms Methodsand Protocols. Springer, USA, Edited byAnton A. Komar, Chapter 14. Methods inMolecular Biology, 578:223-34, 2009.Mismatch Oxidation Assay: Detectionof DNA Mutations using a Standard UV/VIS Microplate Reader. Tania Tabone,Georgina Sallmann, and Richard G.H.Cotton. Single Nucleotide PolymorphismsMethods and Protocols. Springer, USA,Edited by Anton A. Komar, Chapter 15.Methods in Molecular Biology, 578:235-42,2009.MITOMASTER: A Model for Locus-Specific Mutation Databases. Richard G.H.Cotton. In This Issue, Human Mutation,30(1):vii, 2009.A1ATVar: a relational database of humanSERPINA1 gene variants leading toα1-antitrypsin deficiency andapplication of the VariVis software. SophiaZaimidou, Sjozef van Baal, Timothy D.Smith, Konstantinos Mitropoulos, MilaLjujic, Dragica Radojkovic, Richard G.Cotton, George P. Patrinos. HumanMutation, 30(3):308-313, 2009.Clinical Genetics & Human GenomeVariation: The 2008 Human GenomeVariation Society Scientific Meeting.William S. Oetting. Human Mutation,30(5):852-856, 2009.Colección de mutantes. Luis Miguel Ariza.Muy Interesante, September 2009:38-46,2009. (Spanish magazine.)45

Genomic Disorders Research CentreThe unplanned discovery of monoclonalantibodies. Richard G.H. Cotton. TheBiochemist, 31(6), 2009.The Human Variome Project. Richard G.H.Cotton. Opinion, Australian R&D Review,December 2009:11.Collection of variation causing disease:The Human Variome Project. RichardG.H. Cotton. Guest Editorial, HumanGenomics, 3(4):301-3, 2009.During the Past year Professor Cottonhas been invited to speak at anoverwhelming number of internationaland Australian meetings, those he hasbeen able to accept are:Invited Lectureshipsand Chairs (InternationalMeetings):Invited speaker, Establishing a local HVPgroup in Jeddah, Jeddah, Saudi Arabia,March 3.Invited speaker, International symposiumand satellite meeting for new hospitalat Catholic Medical Center/CatholicUniversity of Korea, Seoul, South Korea,April 30-May 1.Invited speaker, Agilent Asia PacificCytogenetics User Summit 2009,Singapore, May 7-8.FutureFest, Cambridge, UK, June 23-25.(Cancelled due to Financial Climate.)Invited speaker, The 3rd Biennial ScientificMeeting, International Society forGastrointestinal Hereditary Tumours, June24-27, Düsseldorf, Germany.Staff ListHead and NHMRC Senior PrincipalResearch FellowRichard Cotton AM BAgSc PhD DSc (Melb)Personal Assistant to the Director/Research AssistantLauren Martin BSc (BiomedSci) (Deakin)BSc (Hons) (Melb)Executive OfficerHeather HowardSenior Research OfficerRania Horaitis BSc (La Trobe)PhD StudentsTimothy Smith BSc (Hons) (Melb)Administrative AssistantAlex Kline BA/BSc (Melb)ConsultantJohn Coghlan AO PhD (Melb)Invited Lectureships AndChairs (Australasian Meetings):Invited Speaker, BioinformaticsSymposium, Genes for Health, TheHuman Genetics Society of Australasia2009 Annual Scientific Meeting,Fremantle, Australia, May 3-6.46

Imaging— Neuroimaging and NeuroinformaticsResearch OverviewUnderstanding the brain requires theintegration of complex neuroscience,and particularly neuroimaging datasets,collected at multiple levels of researchinto brain structure and function. TheNeuroimaging and Neuroinformaticsgroup is at the forefront of rapidlyadvancing interdisciplinary fields thatdevelop and apply imaging techniquesand approaches that are essential toadvancing our understanding of thestructure and function of the humannervous system.Magnetic Resonance Imaging (MRI) is animportant basic and clinical neurosciencediscipline, is able to provide direct in vivomeasurements of the human brain, andis able to provide novel insights intobrain and mind diseases. The researchprojects undertaken in the Neuroimagingand Neuroinformatics Group are aimedat improving our understanding ofneurodegeneration in Friedreich’s ataxia,the identification of brain areas involvedin central control of cough, and thespecialized function of parietal lobe brainregions for performing mental rotationand mental manipulation.Figure 1: Tractography ofsuperior cerebellar peduncle(arrowed) in an individual withFriedreich’s ataxia shows atrophyof the SCP compared with onehealthy control.Major Research ProjectsFriedrich’s AtaxiaNeuroimaging in Friedreich’s AtaxiaDr Hamed Aklaghi, A/Prof NellieGeorgiou-Karistianis (Monash University),Louise Corben (Monash University), ProfElsdon Storey (Monash University), Em.Prof John Bradshaw (Monash University),Dr Andrew Churchyard, Prof MartinDelatycki (Murdoch Children’s ResearchInstitute) and Prof Gary EganFriedreich’s ataxia is a multisystemautosomal-recessive disease, and themost common genetically inheritedataxia affecting approximately 1 in 29,000individuals. FRDA is due to mutations infrataxin gene causing loss of large primaryneurons in the dorsal root ganglia, lossof sensory fibres in peripheral nerves,degeneration of the posterior columnsof the spinal cord and atrophy of thedentate nucleus of the cerebellum.Motor and cognitive dysfunction inFriedreich’s ataxia remains poorlyunderstood. The aims of the study wereto investigate the contributionsof cerebral and cerebellar damagein complex motor and cognitivedysfunction in Friedreich’s ataxia. Wereported that the size of the superiorcerebellar peduncle (the main output ofthe cerebellum from the dentate nucleus)is significantly reduced in Friedreich’sataxia and correlates with age of onset,disease duration and ataxia rating scale.We have also investigated brain whitematter micro-structural changes usingdiffusion tensor imaging (Figure 1).We found that not only the primaryfibres coming out of the dentatenucleus via the superior cerebellarpeduncle (dento-thalamic tract) isaffected, but also post-synaptic fibers(thalamo-cortical tracts) characterisethe same micro-structural changes ofthe primary tract (dento-thalamic tract).We are now evaluating the relationshipsbetween the white-matter changes anddisease parameters.We have investigated the functionaleffects of cerebellar and dentate atrophyand found that whereas in healthy subjectsthe cerebellar motor cortex increasinglyactivates in accordance with thecomplexity of motor tasks, in individualswith Friedreich’s ataxia the cerebellumfunctional activity is unchanged for motortasks of differing complexity (Figure 2).Figure 2: Brain activation during a visuallycued regular finger tapping task, with the blueareas indicating activation in the individualswith Friedreich’s ataxia and red areas showingactivation in control subjects.Figure 2Figure 148

Neural control of coughA/Prof Stuart Mazzone (Universityof Queensland), Dr Leonie Cole,Ms Ayaka Ando, Prof Gary Eganand Dr Michael FarrellExcessive coughing is one of the mostcommon reasons for seeking medicaladvice yet the available therapies fortreating cough disorders are inadequate.We can voluntarily cough, choose tosuppress a cough and we are acutelyaware of an irritation that is present in ourairways. Whilst this indicates a significantlevel of behavioral and conscious controlover the basic cough reflex pathway, verylittle is known about the neural basis forhigher brain regulation of coughing.We used functional brain imaging inhealthy subjects to identify for the firsttime the suprameduallry control ofcough and cough suppression (Figure3). We have shown that the brain regionsactivated during coughing in response toan evoked airways irritation is more thanthe combination of a voluntarily initiatedcough and airways irritation. Rather, brainactivations in the posterior insula andposterior cingulate cortex define theunique attributes of an evoked cough.We have demonstrated for the first timein humans that evoked cough is not solelya brainstem mediated reflex responseto irritation of the airways, but requiresactive facilitation by cortical regions andcan be further regulated by distinct higherorder inhibitory processes.Parietal lobe function in mentalrotation processesDr Andrea Gogos (Mental HealthResearch Institute), Dr Maria Gavrilescu,Dr Sonia Davison (Monash MedicalSchool), Ms Kariss Searle, Ms JennyAdams (MOnash Medical School),Dr Robin Bell (Monash Medical School),Prof Susan Davis (Monash MedicalSchool) and Prof Gary EganMental rotation is a task known to activatethe parietal cortical regions. Our aimwas to investigate whether brain regionswithin the parietal lobe reveal functionalspecialisation with increasing angle ofrotation. Functional MRI was performedwhilst subjects performed a parametricmental rotation task comprising of sixalphanumeric characters presentedin their normal or mirror-reversedorientation. It is well known that subjectsshowed increased reaction times withincreased angle of rotation.Figure 3We observed mental rotation associatedbrain activation bilaterally in the middleoccipital gyrus, left superior temporalgyrus, medial frontal gyrus, and inthe right superior and inferior parietallobules. Brain activation increasedlinearly with increasing angle of rotationto the greatest extent in the superiorparietal lobules compared to the inferiorparietal lobules. Our findings suggestthat posterior parietal lobules performspecialized functions during mentalrotation. A greater understanding ofthe roles of parietal lobe brain regionshelps explain our abilities to mentallymanipulate objects in our “mind’s eye”and may explain our differential abilitiesfor performing mental manipulation skills.Figure 3: Functional brain imaging sectionsshowing activation in suprameduallry regions incough and cough suppression in healthy subjects.49

Imaging— Neuroimaging and NeuroinformaticsOther Research ProjectsThe effects of a noxious coldconditioning stimulus on brainresponses to noxious pressureM.J. Farrell (Centre for NeuroscienceUniversity of Melbourne), T. Tran(Melbourne Health), L.J. Cole, M.Gavrilescu, S.J. Gibson (National AgeingResearch Institute, Melbourne), G.F. EganImproved sensitivity estimation methodfor MR phase image reconstructionZ. Chen, L. Johnston (School ofEngineering, University of Melbourne),N. Faggian, G.F. EganCorpus callosum development in thepreterm infant: a DTI studyD.K. Thompson, T.E. Inder (WashingtonUniversity St Louis, USA), L.A. Johnston(School of Engineering, University ofMelbourne), N. Faggian, L.W. Doyle(Royal Women’s Hospital, University ofMelbourne), G.F. Egan & the VictorianInfant Brain Studies (VIBeS)A stochastic model for characterisingneurodynamic signal and noiseprocesses in fMRIL.A. Johnston (School of Engineering,University of Melbourne), M. Gavrielscu,E.P. Duff, G.F. EganA ranked shape analysis of sub-corticalstructures in Huntington’s diseaseN. Faggian, L.A. Johnston (School ofEngineering, University of Melbourne),I. Bohanna, Z. Chen, N. Georgiou-Karastianis (Monash University), G.F. EganThe optimised projection imagemethod for targeted tissue contrastenhancementA. Ng (Monash University), N. Faggian,Z. Chen, J. Zhang (Monash University),G.F. Egan, L.A. Johnston (School ofEngineering, University of Melbourne)Reduced diffusion anisotropy in opticradiation correlates with optic nerveatrophy after optic neuritisS. Kolbe, C. Bajraszewski, C. Chapman(Royal Eye & Ear Hospital), P. Mitchell(Melbourne Health), L. Johnston (Schoolof Engineering, University of Melbourne),H. Butzkueven, M. Paine (Royal Eye & EarHospital), T. Kilpatrick, G.F. EganThirst activation in the cingulate cortexhas different responses to ad libitumdrinking, water extracted from thestomach after drinking, and input ofwater into the stomachM.J. Farrell, R.E. Shade (SouthwestFoundation, Texas USA), D.A. Denton(Dean’s Office, University of Melbourne),P.T. Fox (Research Imaging Institute,Texas, USA), G.F. EganInvestigating dynamic changes in fMRIfunctional connectivity using timemoving correlationM. Gavrilescu, L.A. Johnston (School ofEngineering, University of Melbourne),M.J. Farrell, G.F. EganFunctional MRI study of Friedreich’sataxia using the Simon TaskH. Akhlaghi, L. Corben (MonashUniversity), E. Storey (Monash University),J. Bradshaw (Monash University), A.Churchyard, N. Georgiou-Karistianis(Monash University), M. Delatycki(Murdoch Children’s Research Institute),G.F. EganBrain responses associated with diffusenoxious inhibitory controls in humans:an ASL fMRI studyL.J. Cole, T. Tran, M. Gavrilescu, S.J.Gibson (National Ageing ResearchInstitute), G.F. Egan, MJ FarrellBaseline axial diffusivity is predictive ofretinal nerve fibre layer thinning afteracute optic neuritisA. Van der Walt, S. Kolbe, A. Klistorner(Sydney Eye Hospital), M. Marriott(Melbourne Health), M. Paine (RoyalVictorian Eye & Ear Hospital), G.F. Egan,Butzkueven, T.J. KilpatrickWhite matter predictors of progressiveneuropsychological and motor deficitsin Huntington’s disease; a DiffusionTensor Imaging studyI. Bohanna, G.F. Egan, J. Stout (MonashUniversity), S-P. Carron, A. Churchyard(Monash University), P. Chua (MonashMedical Center), N. Georgiou-Karistianis(Monash UniversityIncreased iron related MR phasesignals in the striatum in Huntington’sdisease: a novel MR neurodegenerativebiomarkerN. Faggian, N. Georgiou-Karistianis(Monash University), Z. Chen, I. Bohanna,S-P. Carron, L.A. Johnston, J. Stout(Monash University), A. Churchyard(Monash University), G.F. EganExamination of cognitive function inFriedreich ataxia using the Simon taskL.A. Corben, H. Akhalghi, N. Georgiou-Karistianis (Monash University),J.L. Bradshaw (Monash University),E. Storey (Alfred Hospital), G.F. Egan,A.J. Churchyard, M.B. Delatycki (MurdochChildren’s Research Institute)Coil Sensitivity Estimation in PolarCoordinatesK. J. Layton (School of Engineering,University of Melbourne), M. Morelande,Z. Chen, B. Moran (School of Engineering,University of Melbourne), P. M. Farrell,G.F. Egan, L. A. Johnston (School ofEngineering, University of Melbourne)Staff ListFellows, Senior Research Officers andResearch OfficersGary Egan BSc (Hons) PhD (Melb) MBA (Mon)Maria Gavrilescu PhD (Mon)Hong Wang MBBS (Tianjin) MAppSci(Swinburne)Ke Fang BE China PhD (China)(until September, 2009)Zhaolin Chen BEng (HIT) PhD (Monash)Em/Prof Derek Denton MB BS KVA FAAFRACPLeonie Cole BBSc (Hons) (La Trobe)50

Honorary Research OfficersNathan Faggian BDigSys (Hons), PhDMichael Farrell BAppSc (Phty) (Lincoln) MSc(La Trobe) PhD (Melb)Andrea Gogos PhDLeigh Johnston BSc/BE (Hons) PhD (Melb)Neil Killeen BSc (Hons) PhD (ANU)Research Assistantsand Administration StaffTamara-Leigh Brawn BA BSc (Hons) (La Trobe)Grad DipEd (Melb)Mary Joy Gleeson BBus MEdWilson Liu BE Hebei (China) MSSE (Melb)Sarah CarronStudentsHamed Akhlagi MD (Iran)Hamed Asadi MD (Tehran, Iran)India Bohanna BA BSc (Hons) (Mon)Tharushini Bowala BTech (Biomed) (Auckland)Catherine Davey BEE (Hons)Adel Foda (from 07/08)Guangqiang John Geng BE (Beijing) ME(Mon)Susan Illic MBBS (Melb)Scott Kolbe BSc (Hons) MelbParesh Mhaispurkar BSc (Bombay) ME (Mon)(to 08/08)Amanda Ng BEng (Monash) (from 08/08)Deanne Thompson BSc (Melb)William Li MD (Melb)Johnson Yu MD (Melb)Conferences andPresentationsMJ FarrellGetting published, Invited Symposium,Australian Pain Society 29th ASM, Sydney,Australia, April 6th (2009) (Chair)Functional brain imaging of interoception,Invited Symposium, Centre for NationalResearch on Disability and RehabilitationMedicine, University of Queensland,Brisbane, Australia, July 27th (2009)Insights into latent processes usingfunctional brain imaging, AMA Sectionof Medico-Legal Practice, AMA House,Parkville, Australia, October 14th (2009)N KilleenInvited talk at the Victorian eResearchStrategic Initiative Showcase, October(2009)D Xiao, J Ribic, H Butzkueven, K Fang,T Kilpatrick, GF Egan and O SalvadoMethod for optic nerve intensitycharacterization in mice from Mn2 +enhanced MRI images, Proceedings ofSPIE, February (2009)Z Chen, J Zhang, R Yang, P Kellman,LA Johnston and GF Egan2D IIR filter for parallel MagneticResonance image reconstruction,presented at the 7th IEEE InternationalConference on Control & AutomationChristchurch, New Zealand, Nov. (2009)MJ Farrell, T Tran, LJ Cole, M.Gavrilescu, S.J. Gibson and G.F. EganThe effects of a noxious cold conditioningstimulus on brain responses to noxiouspressure, Aust. Pain Society AnnualScientific Meeting, Adelaide (2009)Z Chen, LA Johnston, N Faggianand GF EganImproved sensitivity estimation methodfor MR phase image reconstruction,International Society for MagneticResonance in Medicine, Hawaii,April (2009)DK Thompson, TE Inder, LA Johnston,N Faggian, LW Doyle, GF Egan, and theVictorian Infant Brain Studies (VIBeS)Corpus callosum development in thepreterm infant: a DTI study, InternationalSociety for Magnetic Resonance inMedicine, Hawaii, April (2009)LA Johnston, M Gavrielscu,EP Duff and GF EganA stochastic model for characterisingneurodynamic signal and noise processesin fMRI, International Society for MagneticResonance in Medicine, Hawaii, April(2009)N Faggian, LA Johnston, I Bohanna,Z Chen, N Georgiou-Karastianis andGF EganA ranked shape analysis of sub-corticalstructures in Huntington’s disease,International Society for MagneticResonance in Medicine, Hawaii,April (2009)A Ng, N Faggian, Z Chen, J Zhang,GF Egan and LA JohnstonThe optimised projection image methodfor targeted tissue contrast enhancement,International Society for MagneticResonance in Medicine, Hawaii,April (2009)S Kolbe, C Bajraszewski, C Chapman,P Mitchell, LA Johnston, H Butzkueven,M Paine, T Kilpatrick and GF EganReduced diffusion anisotropy in opticradiation correlates with optic nerveatrophy after optic neuritis, InternationalSociety for Magnetic Resonance inMedicine, Hawaii, April (2009)MJ Farrell, RE Shade, DA Denton,PT Fox and GF EganThirst activation in the cingulate cortexhas different responses to ad libitumdrinking, water extracted from thestomach after drinking, and input ofwater into the stomach, 15th InternationalConference on Human Brain Mapping,San Francisco, June (2009)M Gavrilescu, LA Johnston, MJ Farrelland GF EganInvestigating dynamic changes in fMRIfunctional connectivity using time movingcorrelation, 15th International Conferenceon Human Brain Mapping, San Francisco,June (2009)H Akhlaghi, L Corben, E Storey,J Bradshaw, A Churchyard, N Georgiou-Karistianis, M Delatycki and GF EganFunctional MRI study of Friedreich’s ataxiausing the Simon Task, 15th InternationalConference on Human Brain Mapping,San Francisco, June (2009)51

Imaging— Neuroimaging and NeuroinformaticsLJ Cole, T Tran, M Gavrilescu,SJ Gibson, GF Egan and MJ FarrellBrain responses associated with diffusenoxious inhibitory controls in humans:an ASL fMRI study, 15th InternationalConference on Human Brain Mapping,San Francisco, June (2009)A Van der Walt, S Kolbe, A Klistorner,M Marriott, M Paine, GF Egan,H Butzkueven and TJ KilpatrickBaseline axial diffusivity is predictive ofretinal nerve fibre layer thinning afteracute optic neuritis, World Congresson Treatment and Research in MultipleSclerosis, Montreal, September (2009)I Bohanna, GF Egan, J Stout,S-P Carron, A Churchyard, P Chuaand and N Georgiou-KaristianisWhite matter predictors of progressiveneuropsychological and motor deficitsin Huntington’s disease; a DiffusionTensor Imaging study, World Congressin Huntington’s Disease, Vancouver,September (2009)N Faggian, N Georgiou-Karistianis,Z Chen, I Bohanna, S-P Carron,LA Johnston, J Stout, A Churchyardand GF EganIncreased iron related MR phase signalsin the striatum in Huntington’s disease: anovel MR neurodegenerative biomarker,World Congress in Huntington’s Disease,Vancouver, September (2009)LA Corben, H Akhalghi, N Georgiou-Karistianis, JL Bradshaw, E Storey, GFEgan, AJ Churchyard and MB DelatyckiExamination of cognitive function inFriedreich ataxia using the Simon task,Aust Society Medical Research, Hobart,November (2009)KJ Layton, M Morelande, Z Chen,B Moran, PM Farrell, GF Eganand LA JohnstonCoil Sensitivity Estimation in PolarCoordinates, ISMRM Workshop onParallel Imaging, Santa Cruz, October(2009)JF DomínguezMapping the Brain Mapping Space:The Cognitive Neuroscience of SpatialNavigation and NeuroanthropologicalExtensions for the Study of NavigationalSkills in Nomadic Pastoralists. TheCognitive Specializations of NomadicPastoralism Workshop, Max PlanckInstitute for Social Anthropology, Halle/Saale, Germany, October (2009)A Neuroanthropological Framework forthe Study of the Neural Underpinningsof Culturally Specific Skills. The CognitiveSpecializations of Nomadic PastoralismWorkshop, Max Planck Institute for SocialAnthropology, Halle/Saale, Germany,October (2009)The Neural Basis of CulturalProduction and Reproduction: ANeuroanthropological Perspective.9th Conference of the AustralasianSociety for Cognitive Science,Macquarie University, 30 Septemberto 2 October (2009)Major Collaborative LinksNationalA/Prof Nellie Georgiou-KaristianisDepartment of Psychology,Monash UniversityLongitudinal functional MRI studies ofcognitive decline in pre-symptomaticHuntington’s diseaseDr Caron Chapman and Dr Mark PaineDepartment of Opthamology,Royal Victorian Eye and Ear HospitalMRI investigations in optic neuritispatientsDr Michael DitchfieldHead, MRI, Royal Children’s HospitalHigh field MRI studiesProf Iven MareelsSchool of Engineering,University of MelbourneMagnetic Resonance Imageand signal processingA/Prof Peter FarrellSchool of Engineering,University of MelbourneOptimised MRI acqusitionDr Mark MorelandeSchool of Engineering,University of MelbourneOptimised MRI acqusitionA/Prof Jingxin ZhangMonash UniversityMRI reconstruction and parallel imagingProf Stephen GibsonNational Ageing Research InstituteStudies of central pain processing inageing and Alzheimer’s diseaseProf Lex Doyle and Dr Peter AndersonDepartment of Neonatology,Royal Women’s Hospital andRoyal Children’s Hospital, MelbourneMRI assessment of neonatal braindevelopmentProf David WalkerDepartment of Physiology,Monash UniversityModelling the cortical foldingprocess using MRIDr Stuart MazzoneUniversity of QueenslandImaging the urge to coughDr Ken MatsudaBernard O’Brien Institute of MicrosurgeryQuantiative Peripheral Nerve ImagingDr Nigel Jones, Dr VivianeBouillerent, Howard Ho Fung Tang,Prof Terrence O’BrienDepartment of Medicine – RMH,University of MelbourneMRI investigation of the temporalprogression of epilepsy and anxietypathologies, and the effects ofintervention, in GAERS epileptic ratsDr Sonia Davison, Dr Robin Bell,Prof Susan DaviesDepartment of Medicine, Alfred Hospitaland Monash UniversityAssessing the effects of hormonaltreatments on cognition functionin womenDr Jacqueline OrianLa Trobe UniversityGrey matter pathology with pre-clinicalmurine experimental autoimmuneencephalomyelitis (EAE)52

Prof Ashley Bush andA/Prof David FinkelsteinMental Health Research InstituteManganese enhanced MRI in the APPtransgenic and the ZnT3 knock-outmouse modelsDr Paul Lockhart and Ms Gabi WilsonMurdoch Children’s Research InstituteMRI analyses the absence of Parkinco-regulated gene (Pacrg) occurrenceof hydrocephalus in mouseA/Prof Richard WeisingerLa Trobe UniversityMRI investigation of decreased body fatEditorial PositionsGary EganAssociate Editor, Human Brain Mapping,Wiley PublishersMember, Editorial Board, Neuroimage,Elsevier Publishers, USAMember, Editorial Board, Frontiersin Neuroscience, Frontiers ResearchFoundation, SwitzerlandMember, Editorial Board, Journal ofNeuroinformatics, Humana Press, USAMr Jason LohreyArcitectaNeuroscience InformaticsDr Ann Borda, Dr Steve Melnikoff,Mr Jared Winton and Mr Jason ChenVictorian eResearch Strategic InitiativeNeuroscience InformaticsProf Christopher Rowe, Prof ColinMasters and Prof David AmesAustralian Imaging Biomarkersand Lifestyle ProjectInternationalDr Janniko GeorgiadisDepartment of Anatomy, University ofGroningen Medical Center, NetherlandsNeuronal mechanisms underlying sexualsatiety in menProf Peter Fox and Prof Jack LancasterResearch Imaging Centre,University of Texas, San Antonio, USANeurophysiological mechanismssubserving primal emotionsA/Prof Terri InderWashington University, USANeonatal brain developmentDr Yaou (Euroasia) LiuDepartment of Radiology, XuanWuHospital, Capital Medical University,Beijing, People’s Republic of ChinaProf Zang-Hee ChoNeuroscience Research Institute,Gachon, Korea53

Imaging— MRI DevelopmentOverviewThe Advanced MRI Developmentphysics team at the FNI Austin precincthas established an internationalreputation for innovative methodologicaldevelopment in a number of areas ofmagnetic resonance physics and of MRsignal processing. One of the principalinterests of this team is translationalresearch, with the aim of applying newmethods to important clinical andneuroscientific problems, in particular inthe field of epilepsy (see Major Programin Epilepsy Imaging section). Significantbreakthroughs have been achieved inthe field of diffusion MRI and perfusionMRI, and many ongoing projects aredeveloping these strengths further.Major Research ProjectsDiffusion MRI: How can we measurestructural connectivity in the brain?Donald Tournier, Fernando Calamanteand Alan ConnellyDiffusion MRI is an imaging techniquethat is unique in its ability to probe tissuemicroarchitecture at the cellular levelnon-invasively. It is increasingly used toinvestigate brain white matter and itsdisorders, providing a wealth of importantinformation that cannot be obtained byany other method. White matter consistsof myelinated axonal fibres that connectcortical regions, where the processingof information takes place. Theseconnections are essential for normalbrain function.White matter abnormalities have beenimplicated in many disease states,including multiple sclerosis, stroke,epilepsy, tumours, dementia, and in arange of mental health disorders (egschizophrenia). The pathophysiologicbasis of many of these disorders isthought to be related to abnormalitiesin the structural connections betweendifferent areas of the brain. Furthermore,knowledge of white matter organisationis essential for neurosurgery, to ensurethe preservation of the patient’s mostimportant functions. There is thereforeenormous interest from neuroscientists,psychiatrists and neurosurgeons in usingthis technique.Despite important recent developments,there is currently no reliable methodto properly characterise the structuralconnectivity. One important aspect of theresearch at the BRI is the developmentof improved acquisition and processingmethods for Diffusion MRI, with especialemphasis in its role to study brainconnectivity.Inferring white matter connectivityusing diffusion-weighted imagingDiffusion-weighted MRI is sensitive tothe microscopic motion or diffusion ofwater molecules in the brain along agiven direction. In the white matter, whichconsists of tightly packed bundles ofneuronal axons, the regular arrangementof fibres introduces a directionaldependence of the image intensity (ieanisotropy). This information can beused by the so-called ‘fibre-tracking’algorithms, to track the path of the fibresand to infer the structural connectivity.However, the model currently widely used(known as the diffusion tensor model)is invalid in regions containing multiplewhite matter tracts that cross or pass veryclose to each other. This can lead thefibre-tracking algorithm to venture intoan adjacent tract with very different endpoints, and connections may be inferredthat do not exist in reality.We have developed an alternativeanalysis method to address this limitation,known as the spherical deconvolutiontechnique. We have demonstrated thatthis approach provides fibre orientationdistribution (FOD) estimates that arerobust to the presence of multipleorientations within a voxel, and are notbased on any assumptions regardinghow many fibre orientations are present.The particular spherical deconvolutionapproach that we have developed (knownas constrained spherical deconvolution orCSD) permits the use of superresolution,whereby more FOD parameters areestimated than were actually measured,improving the angular resolution ofthe results. The method provides verywell defined fibre orientation estimates(Figure 1), which enable tractographyalgorithms to be able to track reliablythrough crossing fibre regions. Usedin conjunction with advanced trackingalgorithms, in particular those whichadopt a probabilistic approach, theCSD method can generate either wholebrain fibre maps (Figure 2) or imagesof individual fibre bundles of interest(Figure 3). By minimising the confoundingeffects of crossing fibres, the CSDmethod provides an important tool forneuroscientific and clinical investigationsof brain connectivity. During 2009, thesoftware that we developed both toperform spherical deconvolution (SD),and to perform in vivo white matter fibretracking using the fibre orientationsgenerated by SD, has been available viathe BRI website as a software package(MRtrix) that has been downloadedwidely by researchers working in thisfield internationally.54

Figure 1Figure 2Figure 3Figure 1: Left: coronal structural MR image ofthe brain. Yellow box indicates a region known tohave complex white matter fibre crossings. Right:Fibre orientation distributions (in region shown onthe left image) determined using our constrainedspherical deconvolution (CSD) technique.Figure 2: Left: Whole brain fibre map resultingfrom application of a probabilistic afibre trackingalgorithm to fibre orientation informationgenerated using the CSD method. Right: 2mmcoronal section from the whole brain trackingdataset, clearly indicating the widespreadpresence of crossing fibre regions (where thediffusion tensor model is inadequate).Figure 3: Images of individual fibre bundles ofinterest. Left: cortico-spinal tract. Right: opticradiation) generated using CSD + probabilistictracking together with tract editing techniques(ie display only tracts that pass through both aseed region and a target region).Perfusion MRI: How can we measurecerebral blood supplyFernando Calamante, Lisa Willatsand Alan ConnellyPerfusion MRI is a non-invasive imagingtechnique for measuring cerebralperfusion (blood delivery to brain tissueper unit time). Blood delivers oxygenand nutrients to the tissue, which arenecessary for cellular metabolism.The survival of the brain is dependenton a continuous and adequate supplyof blood, and failure of the cerebralcirculation can result in cell death.Similarly, some clinical conditions areassociated with a hyperperfusion status(such as epilepsy and tumours) due totheir increased energy demand. For thesereasons, the ability to measure perfusionaccurately, noninvasively, and with goodspatial resolution would offer the chanceto identify and characterise abnormaltissue in many clinical conditions.A significant expansion in the availabilityof MRI scanners has taken place in the lastdecade, and Perfusion MRI has becomean important diagnostic technique. Everymajor MRI scanner manufacturer providesimaging sequences for Perfusion MRI,but their product analytical software isstill somewhat rudimentary. Improvedmethods to quantify perfusion andcharacterise the vascular networks inthe brain is an important aspect ofresearch at the BRI.Brain perfusion quantificationin patients with strokeBolus-tracking MRI is the most commonlyused Perfusion MRI technique in clinicalstudies. It involves injection of a bolusof contrast agent in the arm, which canbe used to measure blood flow in thebrain. Perfusion quantification requiresmeasurement of the arterial inputfunction (AIF), which describes the inputof contrast agent to the tissue. Althoughthis function can vary throughout thebrain, a single (ie global) AIF is commonlyused in practice. However, the presenceof vascular abnormalities (eg arterialstenosis, occlusion, or collateral supply)may cause distortions in the bolus (delayand dispersion), which we have showncan lead to severe local underestimationsin perfusion quantification.55

Imaging— MRI DevelopmentOne solution to this delay and dispersionproblem involves estimation of alocal AIF from a small vessel closer tothe tissue of interest. However, this isproblematic due to partial volume effects(ie contamination with the surroundingtissue). We have recently developeda method to define a local AIF usingindependent component analysis, atechnique designed to identify temporal/spatial independent patterns. Using thismethodology, the underestimation inperfusion quantification is minimised(Figure 4).What is the blood flow pattern in thebrain vasculature?Another important aspect ofunderstanding cerebral perfusion isto determine the arterial origin forthe blood supplied to the differentregions of the brain, ie the vascularnetwork. This information, known as thecerebral arterial territories, can play akey role for the management of strokepatients in the differential diagnosis ofhaemodynamic border zone infarctionand thromboembolic ischemia.However, individual arterial territories aredifficult to identify in vivo, interpretationof acute stroke subtype and underlyingpathogenesis, based on the topographicpatterns alone, is often inaccurate.Therefore, there is a need for thedevelopment of new techniques forthe mapping of arterial territories. Thecurrently limited understanding ofthe collateral circulation in the humanbrain can be greatly enhanced throughthe development of vascular models,providing the basis for future therapeuticand prognostic applications.As part of a collaboration with Dr J.R.Cebral (George Mason University, USA)and Dr Roland Bammer (StanfordUniversity, USA), we have developed anovel method to create a subject-specificvascular model, which is based oncombining high-resolution MR imagesand MR angiography with computationalfluid dynamic modelling (Figure 5). Thesevascular models can greatly enhancethe current limited understanding of thecollateral circulation in the human brain,providing the basis for future therapeuticand prognostic applications.Figure 4Figure 5Figure 4: In patients with stroke, mismatchbetween lesion size on diffusion weighted MRI(DWI) and that indicated by perfusion MRImeasures (eg mean transit time (MTT)) is believedto identify tissue that is under threat of infarction.Decision to treat a patient with thrombolytictherapy may be based on such information.This example shows a small acute DWI lesion(left), extensive perfusion deficit (increased MTT,coloured region on MTT maps) using global AIF,and small extent of perfusion deficit using a localAIF. The large mismatch area using the globalAIF would suggest the need to treat, while thefollow-up final infarct size indicates that this isincorrect. The local AIF more correctly indicatedminimal mismatch area. Since thrombolysis hasan associated risk of haemorrhage, it is essentialthat perfusion MRI data are accurate if they are toguide patient selection.Figure 5: Top: Visualizations of the blood-flowpatterns (represented as streamlines) at peaksystole in the major cerebral arteries in a normalsubject obtained from MRI (phase-contrast MRI)data and a Computational Fluid Dynamics (CFD)model. Bottom: Wall shear-stress distributions inthe same subject determined from CFD models.56

Other Research ProjectsDevelopment of a new fibre trackingalgorithm for visualising white matterfibre tracts in the brainAlan Connelly, Fernando Calamante,Donald Tournier (BRI), Leigh Johnston,Iven Mareels (Electrical Engineering,University of Melbourne) andThomas Close (PhD student,University of Melbourne)In order to undertake this researchproject, a joint PhD position has beenestablished between FNI and the Dept ofElectrical Engineering at the University ofMelbourne. The overall aim is to developa novel approach to fibre tracking in thebrain in vivo; since there is currently norecognised gold standard by which tojudge the success of any such algorithm,the project started by undertakingthe development of a software toolto generate simulated white matterstructures. Since the configuration of thesimulated fibres is known, this tool willenable the testing of novel algorithmsfor performing tracking of brain whitematter in vivo. This work was publishedin the journal NeuroImage in 2009. Asoftware package has also been madeavailable via the Brain Research Institutewebsite to enable the tool to be usedby other researchers in the internationalacademic community. Continuation ofthe project has made significant progresstowards the development of a new fibretracking algorithm that can incorporateinformation from remote voxels to informthe estimation of fibre orientation withinany individual voxel, ie by neighbourhoodtrack modelling.Automatic clustering of fibre tracksin whole brain fibre tracking data setsAlan Connelly, Fernando Calamante,Donald Tournier (FNI) and Robert Smith(PhD student)Whole brain tracking datasets (consistingof 1 million tracks or more) present asignificant problem in terms of their size,to the extent that no current fibre-trackclustering algorithm can come close todealing with such large amounts of data.It is desirable therefore to develop amethod by which such large datasets canbe clustered into biologically meaningfulfibre bundles. We are undertaking tofind a solution to this problem, capableof automated clustering of very largeprobabilistic track data sets at anychosen cluster scale. This would providean invaluable tool for the investigationof structural connectivity within theabnormal brain.Replacement for FA in voxelscontaining crossing fibres (>90%of white matter voxels in brain)Alan Connelly, Donald Tournier (FNI),Olivier Salvado (CSIRO, Brisbane),Stuart Crozier (University of Queensland)and David Raffelt (PhD student, UQ)Numerous neuroscientific studies haveused voxel based analysis of diffusiontensor (DT) invariants, such as fractionalanisotropy (FA), to investigate groupdifferences in white matter. However,results are difficult to interpret in regionswith crossing fibres due to the simpleGaussian diffusion model assumedwith the DT that is inappropriate insuch regions. High Angular ResolutionDiffusion-weighted Imaging (HARDI) hasbeen employed by a number of higherorder models (HOM) (such as Q-ball andour own method, Constrained SphericalDeconvolution (CSD)) to resolve crossingfibres by estimating diffusion or FibreOrientation Distribution (FOD) functions.Fibre tractography and connectivitystudies benefit from HOM, howeverHOM for whole brain voxel basedanalysis has been largely unexplored.We are developing a novel measure toenable voxel wise comparisons to bemade over all orientations.Improved methods to enable analysesof high angular resolution DW imagingdata across large groups of subjectsAlan Connelly, Donald Tournier (FNI),Olivier Salvado (CSIRO, Brisbane),Stuart Crozier (University of Queensland)and David Raffelt (PhD student, UQ)In order to use diffusion weightedMRI data to investigate differencesbetween patient populations andnormal subjects, it is essential to havegood spatial registration betweensubjects. In this study, we are developinga registration method using a novelsimilarity measure that optimises directlyon Fibre Orientation Distributions (FOD).Unlike existing techniques, this exploitsthe additional information providedin voxels with crossing fibres, and isindependent of the model used (egQ-Ball or CSD). More accurate spatialnormalisation will benefit group basedclinical and neuroscientific studies, aswell as automatic atlas based anatomicallabelling, which are increasingly importantapplication areas for diffusion MRI.Identifying brain damage underlyingspeech deficits in children followingtraumatic brain injury: A new modelof diagnosis and prognosisAlan Connelly, Donald Tournier (FNI),Angela Morgan, Sheena Reilly, VickiAnderson (MCRI) and Frederique Liegeois(University College London)Our aim is to link state-of-the-art MRIfindings to quantitative speech datato: determine the association betweenregions of brain damage and speechdisorder-post TBI; examine neuralmechanisms controlling speech prognosisby determining regions of brain damageassociated with persistent vs recoveredspeech; and develop a world-first speechdiagnostic system for children withspeech disorder post-TBI.Physiological Significance of theTime-to-Maximum (Tmax) Parameterin Perfusion MRIFernando Calamante, Alan Connelly, (FNI),Søren Christensen, Patricia DesmondStephen Davis,(Royal Melbourne Hospital),and Leif Østergaard (Århus UniversityHospital, Denmark)Many perfusion-related MRI parametersare used to investigate the so-calledpenumbra in stroke (ie a region ofhypoperfused tissue surrounding aninfarct core that is at risk of proceedingto infarction). Although time-to-maximum(Tmax) of the residue function hasbeen suggested as a very promisingparameter, its physiological meaning andsensitivity to experimental conditionsare not well-understood. This studywill use simulations to further ourunderstanding of the practical meaningof Tmax, including its dependence onsuch things as delay, dispersion, andmean transit time. Our aim is to providerecommendations for its use in clinicalinvestigations through an investigationof in vivo examples guided by thesimulation findings.57

Imaging— MRI DevelopmentImproved Manual and AutomaticSelection of the Arterial Input Functionin Dynamic Susceptibility Contrast MRIFernando Calamante, Alan Connelly(FNI), Egbert Bleeker, Matthias van Osch,Mark van Buchem and Andrew Webb(Leiden University).Dynamic susceptibility contrast-MRIrequires an arterial input function (AIF)to obtain cerebral blood flow, cerebralblood volume, and mean transit time. Thecurrent AIF selection criteria discriminatevenous, capillary, and arterial profilesbased on shape and timing characteristicsof the first passage. Unfortunately, partialvolume effects can lead to shape errors inthe bolus passage, including a narrowerand higher peak, which might be selectedas a ‘‘correct’’ AIF. In this study, a newcriterion is being investigated to detectshape errors based on tracer kineticprinciples for computing cerebral bloodvolume. This criterion will make use ofthe ratio of the steady-state value to thearea-under-the-curve of the first passage,which should result in an equal value fortissue and arterial responses. By usinga reference value from tissue, partialvolume effects–induced shape errors ofthe AIF measurement should be able tobe detected.Staff and StudentsProf Alan Connelly Head of Advanced MRIDevelopmentDr Fernando Calamante Senior Lecturer inMR PhysicsDr Donald Tournier Post Doctoral PhysicistDr Lisa Willats Post Doctoral PhysicistDr Xiaoyun Liang Post Doctoral PhysicistThomas Close PhD studentDavid Raffelt PhD studentElizabeth Smith PhD studentRobert Smith PhD studentDr Mark MacKay PhD studentDr Susan Palmer Research AssistantGagan Sharma Research AssistantInvited Oral PresentationsAlan ConnellyStructural and diffusion MRI EducationalCourse, 15th Annual Meeting of theOrganisation for Human Brain Mapping,San Francisco, USANeuroscience Lecture Series, FloreyNeuroscience Institutes, Melbourne,AustraliaComputational Neuroscience Symposium,Melbourne, AustraliaFernando CalamanteNeuroscience Symposium, Melbourne,AustraliaFlorey Neuroscience Institutes (FNI)Parkville Seminar Series, Melbourne,AustraliaEpilepsy Research Retreat, HepburnSprings, AustraliaClinical Neuroscience Research &Neuroimaging Course, Melbourne,AustraliaWeekend Educational Course in theInternational Society for MagneticResonance in Medicince Annual Meeting,Honolulu, Hawaii, USAJ-Donald TournierNeonatology Unit, Karolinska Institute,Stockholm, SwedenUCL Insitute of Child Health, UniversityCollege London, London, UKShort course in Diffusion Imaging, Princeof Wales Medical Research Institute,Sydney, NSWMelbourne University NeuropsychologyStudent’s Society, Melbourne, AustraliaCentre for Clinical Research, University ofQueensland, Brisbane, AustraliaQueensland Cerebral Palsy &Rehabilitation Research Centre, Universityof Queensland, Brisbane, AustraliaThree hour-long presentations at theClinical Neuroscience Research &Neuroimaging Course. Melbourne,AustraliaWhite matter tractography using diffusionMRI. Neuroscience seminar series,Royal Melbourne Hospital, Melbourne,AustraliaCrossing fibers in Diffusion MRIEducational Course.16th annual meetingof the International Society for MagneticResonance in Medicine, Honolulu,Hawai’i, USANational and InternationalCollaborative LinksProf Sam.Berkovic and Prof IngridScheffer (University of Melbourne),Prof Graeme Jackson and Dr StevePetrou (Florey Neuroscience Institutes),Prof David Reutens (University ofQueensland), Prof John Mulley andProf Josef Gecz (University of Adelaide)As part of a NHMRC Program Grant team,the investigation of the molecular basisfor and brain abnormalities associatedwith a range of epilepsiesProf Leif OstergaardCenter for Functionally IntegrativeNeuroscience, Århus University Hospital,DenmarkImproved methods to measure perfusionusing MRIA/Prof Matthias van OschLeiden University Medical Centre, Leiden,The NetherlandsInvestigation of contrast-agentquantification issues in perfusion MRIA/Prof Juan CebralGeorge Mason University, Virginia, USAMRI data computational fluid dynamicsmodelling to develop models of vascularterritories and collateral circulation ofthe brainDr Roland BammerStanford University, USAComparison of MRI measurementsof blood flow in the arteries withthose predicted using computationalfluid dynamicsDr David ThomasUniversity of London, UKDevelopment of improved method ofquantifying cerebral blood flow usingarterial spin labelling58

Dr Chin-Po LinNational Yang-Ming University,Taipei, TaiwanTo investigate MRI methods to identifywhite matter fibre orientations in thebrain using experimental water phantomsProf Geoffrey A. Donnan and A/ProfHelen Dewey, NSRICharacterisation of the penumbra inpatients with acute stroke using advancedperfusion MRI techniquesDr Amy Brodtmann, NSRITo establish whether arterial stenoses cancause cortical remodelling within brainregions supplied by the affected vesselProf Sheena Reilly , Dr Angela MorganMurdoch Children’s Research InstituteTo determine the structural-functionalabnormalities in children withdevelopmental language disordersDr Mark MackayMurdoch Children’s Research InstituteCharacterisation of the ischaemicpenumbra in children with strokeProf Stephen Davis, Dr PatriciaDesmond, Dr Soren ChristensenRoyal Melbourne HospitalTo develop improved perfusion MRImethods for the analysis of the MRperfusion data from patients with strokeProf Iven Mareels, Dr Leigh JohnstonDepartment of Electrical and ElectronicEngineering, University of MelbourneDevelopment of improved methods ofmeasuring white matter connectivityusing diffusion MRIA/Prof Gavin FabinyiAustin HealthInvestigation of the efficacy of diffusionbased MRI white matter fibre trackingin predicting the visual field deficitscommonly experienced by patientsundergoing temporal lobe neurosurgeryDr Torsten Baldeweg, Dr FrederiqueLiegeois, Dr Brigitte VollmerInstitute of Child Heath, UniversityCollege London and Karolinska Institute,Stockholm, SwedenCombined neuropsychological andneuroimaging study of speech andlanguage function in preterm childrenwith perinatally acquired brain lesionsDr Martin King, Dr Brigitte Vollmer,Dr Samuel GroeschelInstitute of Child Health, UniversityCollege London, and Karolinska Institute,Stockholm, SwedenNeuroimaging study of normal braindevelopment from infancy to adulthoodProf David Gadian, Dr WK Chong,Dr Frederique Liegeois andProf Helen CrossInstitute of Child Heath, UniversityCollege LondonFunctional neuroimaging studies inchildren with focal epilepsyProf Lex Doyle, Dr Peter Anderson,Prof Terrie Inder, Prof Jeff NeilRoyal Women’s Hospital, Melbourneand St Louis, USADetermining the mechanisms leadingto long term impairment in verypre-term childrenDr Olivier Salvado, Prof Stuart CrozierCSIRO, University of QueenslandTo develop methods to spatially normalisewhite matter fibre orientation imagesacross subjects and perform groupanalyses of high angular resolutiondiffusion weighted MRI dataEditorial PositionsProf Alan ConnellyEpilepsiaGrantsS.F.Berkovic, G.D.Jackson, J.C.Mulley,D.C.Reutens, S.Petrou, I.E. Scheffer,J.Gecz, and A.ConnellyEpilepsy: Molecular basis andmechanisms in the era of functionalgenomics. National Health and MedicalResearch Council. (Program Grant). 5years (2006-2010). $12,015,768A.Morgan, A.Connelly, and F.Liegeois.Identifying brain damage underlyingspeech deficits in children followingtraumatic brain injury: A new modelof diagnosis and prognosis. VictoriaNeurotrauma Initiative. 3 years (2008-2010). $277,000.F.Calamante, A.Connelly, and H DeweyImproved identification of at risk braintissue in patients with stroke. NationalHealth and Medical Research Council(Project Grant). 3 years (2008-2010).$277,276G.Donnan, S.Davis, and A.ConnellyStroke Imaging Prevention and Treatment.CSIRO Flagship Collaboration ClusterFund. 3 years (2008-2011). $3,000,000.D.F.Abbott, G.D.Jackson, andA.ConnellyA device for simultaneous continousacquisition of EEG and MRI. NationalHealth and Medical Research Council(Development Grant). 1 year (2008-2009).$179,401.F.CalamanteNational Health and Medical ResearchCouncil Career Development Award.5 years (2007-2011), $445,00059

Imaging— MRI DevelopmentPublications2009 Peer reviewed publicationsCalamante F, Connelly A. PerfusionPrecision in Bolus-Tracking MRI:Estimation Using the Wild-BootstrapMethod. Magnetic Resonance inMedicine 2009; 61:696-704. 2009Calamante F, Connelly A, van Osch MJP.Nonlinear Delta R-2* Effects in PerfusionQuantification Using Bolus-Tracking MRI.Magnetic Resonance in Medicine 2009;61:486-492. 2009Close TG, Tournier JD, Calamante F,Johnston LA, Mareels I, Connelly A. Asoftware tool to generate simulated whitematter structures for the assessment offibre-tracking algorithms. NeuroImage 47:1288-1300. 2009De Tiege X, Connelly A, LiegeoisF, Harkness W, Clark CA, Chong WK,Gadian DG, Cross JH. Influence of motorfunctional magnetic resonance imagingon the surgical management of childrenand adolescents with symptomatic focalepilepsy. Neurosurgery 2009;64(5):856-64.2009O’Reilly M, Bollmer B, Vargha-KhademF, Neville BGR, Connelly A, Wyatt J,Timms C, de Haan M. Ophthalmological,cognitive, electrophysiological andMRI assessment of visual processingin preterm children without majorneuromotor impairment. DevelopmentalScience 2009; 1-14. 2009Cebral JR, Putman CM, Alley MT, Hope T,Bammer R, Calamante F. Hemodynamicsin Normal Cerebral Arteries: QualitativeComparison of 4D Phase-ContrastMagnetic Resonance and Image-BasedComputational Fluid Dynamics. J. Eng.Math. 64:367–378. 2009Book chapters – in pressF. Calamante, J-D.Tournier and A.Connelly.Stroke and cerebrovascular diseasein childhood: Recent advances in MRimaging. In: Stroke and cerebrovasculardisease in childhood. (Editors: GanesanV and Kirkham FJ). MacKeith Press(Available from Feb 2010). ISBN:9781898683346.J-D Tournier. Diffusion MRI: Crossingfibres in practice. In: Diffusion MRI (Editor:Derek Jones). Oxford University Press.(Available from May 2010).60

Multiple SclerosisResearch OverviewMultiple sclerosis (MS) is the mostcommon neurodegenerative diseasethat affects young adults in ourcommunity and is widely considered tobe an enigmatic disease that involvesautoimmune mechanisms, but whoseseverity is dependent upon the natureof the neurobiological response toinjury. Our division strives to understandthe cause of MS and to develop bettertherapies to treat the condition. Toachieve these aims, we are focusingon several areas, including the geneticdeterminants of MS, the potentialof neuroprotective and regenerativemedicine and the development of novelprognostic indicators.Our genetics research has identifiedtwo novel loci that confer susceptibilityto multiple sclerosis. One of these lociappears to influence the expressionof the CD40 protein, which modulateskey aspects of immune function andwhich has also been implicated in thepathogenesis of other autoimmunediseases, notably Grave’s disease andrheumatoid arthritis. The other locusencodes a region of the genome thatencompasses some 17 genes, includingkey candidate genes involved in VitaminD metabolism, which we and othershave previously shown to be potentiallyimportant in the susceptibility to MS.Our focus on regenerative medicinefollows on from seminal work performedsome years ago which we contributedto, that identified stem cell populationswithin discrete regions of the adultmammalian brain, in particular thesubventricular zone (SVZ). By studyingthe molecular response of the SVZ to ademyelinative insult, we have identifiedcandidate genes that are potentiallyimportant in modulating regeneration.One important set of candidates is thebone morphogenic protein family andwe are currently exploring the influenceof this series of factors on precursor cellsand their progeny during demyelination.Another area of interest is to developstrategies focusing on cellular protection,designed to potentiate the survivalof either the oligodendrocyte, themyelinating cell of the central nervoussystem or neurons. We have a strongtradition in this area, having previouslyidentified that molecular signalinginduced by the growth factor, LIF, is a keycomponent of the protective responseinitiated within oligodendrocytes, asa consequence of a demyelinativeinsult in vivo. Our more recent workhas identified that the TAM family ofreceptor tyrosine kinases and their ligandGas6 are also important in promotingoligodendrocyte survival but also inmodulating innate immune activityduring central demyelination.We are also developing biomarkersof disease severity and continue to beencouraged by work that has shownthat release into the serum of theneurofilament heavy chain from damagedneurons could be a reliable and validmarker of ongoing neuronal damage.Such a marker should prove invaluable infuture clinical studies designed to assessthe efficacy of novel neuroprotective andregenerative therapies in MS.Major Research ProjectsRole of TAM receptor signalling incentral nervous system demyelinationMichele Binder, Holly Cate, Junhua Xiao,Dennis Kemper, Gerry Ma, Judith Field,Melissa Gresle, Simon Murray, HelmutButzkueven, Catriona McLean andTrevor J KilpatrickThe TAM family (Tyro3, Axl and Mer) ofprotein tyrosine kinase receptors playpivotal roles in the processes of cellsurvival and proliferation, modulation ofthe immune response, and the removalof dead cells from tissue. Disruption ofthese processes has been shown to becentral to both the initial development,and the subsequent clinical course,of demyelinating diseases. All threereceptors, and their ligands Gas6 andProtein S, are expressed in the centralnervous system (CNS), including inoligodendrocytes, the myelin-producingcell of the CNS.Previous work from our laboratory hasshown that loss of Gas6-dependant TAMreceptor signalling negatively affectsthe survival of oligodendrocytes duringcuprizone-challenge, an experimentalmouse model of demyelination. In thismodel, the loss of Gas6, and subsequentdecrease in oligodendrocyte survival,was accompanied by an increase inthe response of the microglia, theinnate immune cell of the central nervoussystem.In addition to regulating the responsesduring demyelination, TAM receptorsignalling also appears to be involvedin the repair response that followsdemyelination. We found that in theabsence of Gas6, whilst initial recoverymatched that of mice with functionalGas6, continued recovery was slowed,although not ultimately prevented. Tounderstand the molecular mechanismsthat drive the observed effects, weexamined the effect of exogenousGas6 in in vitro myelination assays. Wefound that Gas6 significantly increasedmyelination in a dose-dependent manner,suggesting that TAM receptor signallingcould be directly involved in myelinationby oligodendrocytes. The reduced rateof remyelination of Gas6 KO mice couldthus result from a lack of Gas6 at a criticaltime during myelin production after injury.Spastic paralysis precedes overtdemyelination in a transgenic model ofinducible oligodendrocyte apoptosisLaura Oluich, Pikying Soo, Sze Woei Ng,Holly Cate, Trevor J Kilpatrick andTobias MersonOligodendrocyte (OL) apoptosis isamong the earliest histopathologicalfeatures of multiple sclerosis (MS). Tomodel OL apoptosis in vivo, we havegenerated transgenic mice in whichOLs are rendered selectively sensitiveto Diphtheria toxin (DT)-mediatedapoptosis. MBP-DTR transgenic miceexpressing Diphtheria toxin receptor(DTR) under the regulatory control of thetruncated mouse myelin basic protein(MBP) promoter exhibit OL-specific DTRexpression in the CNS. Administrationof 200ng DT to MBP-DTR mice resultedin the development of a progressiveneurological phenotype characterisedby spastic paralysis of the hind limbs.Histopathological analysis of DTchallengedMBP-DTR mice at the peak ofclinical disease revealed a 35% reductionin the density of CC1-immunoreactiveOLs relative to DT-challenged wild typemice (n=4 per genotype). Despite lossof OL cell bodies, myelin integrity, asassessed by RIP immunohistochemistry,was unaffected. Although myelinappeared to be grossly normal inclinically affected DT-challenged MBP-DTR mice, the presence of b-amyloid62

precursor protein-immunoreactiveaxonal swellings suggests that axonalprotein trafficking was disrupted insome neurons. Furthermore, a markedinflammatory response was indicated bythe presence of numerous activated Iba1-immunoreactive microglia, particularlyin grey matter. Our results suggestthat neuronal pathology following OLapoptosis precedes demyelination andimplies that microglia could be prephagocyticat this time-point. We arecurrently examining myelin integrity atthe ultrastructural level and characterisingchanges in the expression of ion channelsand adhesion proteins at paranodal andnodal regions of the axo-glial junctionsof the MBP-DTR mice. Ongoing analysesusing the MBP-DTR model will providea unique opportunity to characterise themolecular and cellular consequencesof OL apoptosis, a hallmark of newlyformingMS lesions, and for defining newstrategies to facilitate neuroprotectionand repair.Glycoprotein 130 signalling regulatesastrocyte production but not selfrenewal of adult neural precursor cellsTobias Merson, Davina Mareels, PikyingSoo, Sze Woei Ng, and Trevor J. KilpatrickCytokines that belong to the IL-6 familyhave been implicated in the maintenanceof various stem cell populations. In theCNS, LIF has been proposed to regulatethe maintenance of neural precursorcells (NPCs) in the embryonic and adultCNS. LIF signals via a heterodimericreceptor complex comprising LIFRb andgp130, the latter being the commonreceptor component for all IL-6 familycytokines. We assessed the consequenceof conditional deletion of gp130 uponthe clonogenicity of NPCs derived fromthe subventricular zone (SVZ) of adultmice. We found that gp130-deficiencyhad no influence upon the capacityof primary neurosphere cells to formsecondary neurospheres, nor did it alterthe growth characteristics of long-termserially-passaged neurosphere cultures.Consistent with this finding, we foundthat stimulating wild-type primaryneurospheres with LIF did not modulatetheir capacity to generate secondaryneurospheres, irrespective of whetherEGF or FGF-2 was used, the duration ofexposure or concentration of exogenousLIF. Under differentiating conditions,gp130-deficient NPCs exhibited amarked deficiency in their capacity togenerate astrocytes of which the vastmajority exhibited low-level expressionof GFAP but normal levels of S100b.Assessment of the effect of exogenousLIF upon the differentiation of wild-typeneurospheres corroborated this result,demonstrating that LIF acted specificallyduring NPC differentiation, rather thanthe proliferative phase, to increase theproduction of astrocytes 2.5-fold relativeto untreated cultures. Together, ourresults suggest that rather than regulatingthe adult NPCs self-renewal, gp130signalling acts to selectively regulatethe production of astrocytes duringNPC differentiation.The role of disabled-2 in murineexperimental autoimmuneencephalomyelitisVilija Jokubaitis, Tania Cipriani,Dennis Kemper, Helmut Butzkuevenand Trevor J KilpatrickNeuroinflammation regulates bothdisease pathogenesis and repairin multiple sclerosis (MS) and ismodeled in experimental autoimmuneencephalomyelitis. Using the EAE model,we identified genes that are differentiallyexpressed by glia in the context ofneuroinflammtion relative to healthand in particular established that theintracellular adaptor protein disabled-2(Dab2) was up-regulated in the diseasestate an average of 3.4 ± 0.7 fold. Weestablished that dab2 gene expressionwas positively correlated to EAE diseaseseverity (r=0.92, p

Multiple SclerosisFigure 1Increasing oligodendrocyte numberswithin myelin lesions by modulation ofbone morphogenic protein signallingJennifer Sabo, Daniel Merlo, Trevor J.Kilpatrick and Holly S. CateEnhancement of endogenousoligodendrocyte regeneration is apromising strategy for repair in chronicdemyelinating diseases of the centralnervous system. Myelin injury inducesseveral factors, including thosethat could either enhance or inhibitoligodendrogliogenesis. We havepreviously shown that bone morphogenicprotein (BMP) signalling is increased inmyelin lesions during cuprizone-induceddemyelination and that BMP4 inhibitsproduction of oligodendrocytes in cellculture. More recently, we have examinedthe effects of modulating BMP signallingon proliferation and differentiation ofadult oligodendrocyte progenitor cells(OPCs) within myelin lesions in the mousebrain. We used osmotic mini-pumps toinfuse BMP4, its endogenous antagonistNoggin or vehicle into the lateral ventricleof the brain during cuprizone-induceddemyelination. We first confirmed thatinfusion of BMP4 significantly increasedBMP signalling activity in the midlinecorpus callosum while Noggin infusionsignificantly decreased BMP signallingactivity. When the tissue was assessedduring demyelination, BMP4 infusedmice had increases in proliferation andoligodendrocyte progenitor cells in themidline corpus callosum compared tovehicle infused mice, while there wasno significant difference in Noggininfused mice. However, when the tissuewas assessed after 1 week of recoveryfollowing demyelination, Noggin infusedmice had increases in the number ofmature oligodendrocytes in the midlinecorpus callosum compared to vehicleinfused mice, while there was nosignificant difference in BMP4 infusedmice. Thus, although BMP infusionresulted in increased OPC proliferationduring demyelination, our results suggestthat Noggin infusion may have a morebeneficial effect on repair given thatNoggin, and not BMP4, infusion resultedin an increase in oligodendrocyte cellnumbers during recovery. These findingssuggest that modulating BMP signallingduring demyelination alters proliferationand differentiation of adult OPCs. Weare currently examining how modulationof BMP signalling affects the astroglialand microglial populations within themyelin lesion and the ultimate effecton remyelination.Validation of PhosphorylatedNeurofilament-H as a serum markerof neurodegeneration in MultipleSclerosis PatientsMel Gresle, Trevor J Kilpatrick and HelmutButzkueven in conjunction with G. Shawof the University of Florida, Y Liu ofXuanwu Hospital Beijing, M. Marriott ofthe Royal Melbourne Hospital, B Taylorof the University of Tasmania andO Skibina, K-J Lazarus and J Haartsenof Box Hill HospitalIn multiple sclerosis (MS), progressiveaxonal loss occurs from disease onset andis thought to be the main pathologicaldeterminant of permanent neurologicaldisability. Although it is clear that novelneuroprotective therapies are required forthe treatment of MS, their developmenthas been hindered by a lack of specificand sensitive measures of axonal injuryin humans, which are needed to monitordisease activity and to compare theefficacy of putative neuroprotectivetherapies. To address this need, we havedeveloped a novel ELISA method formeasuring phosphorylated neurofilamentH (pNF-H) protein, which is only found inaxons. When axons are damaged, pNF-His released and moves into the bloodwhere it can be measured to indicate howmuch damage has occurred. We havealready shown that this ELISA method canbe used to measure axonal damage in amouse model of MS, and we would nowlike to determine whether this assay islikely to be useful for monitoring axonalinjury in MS patients.Thus far we have measured serum pNF-Hlevels for 59 RR-MS and 136 healthycontrols, and have found that 15% of RR-MS patients have clearly elevated serumlevels of this protein, relative to healthycontrol levels. These patients also havean average disability score that is 2 pointshigher than patients with normal serumlevels of this protein. These preliminarystudies provide strong evidence thatelevated serum pNF-H levels aredetectable in a proportion of patientswith RR-MS, using the ELISA methodrefined in our laboratory in collaborationwith its inventor, Professor Gerry Shaw.Importantly, serum pNF-H levels arehigher in patients with more rapidlyprogressing disease, suggesting it couldbe an indicator of rapidly accumulatingaxonal injury in MS patients. If we areable to confirm these observations, thisassay could provide an important toolfor testing the effectiveness of novelneuroprotective therapies for MS; orit could be used to improve diseasemonitoring in MS patients, by indicatinghow they are responding to therapies.The role of the neurotrophins inregulating myelinationJunhua Xiao, Melanie Willingham,Agnes Wong, Trevor J Kilpatrickand Simon MurrayThe group is currently characterizing theinfluence that the neurotrophin BrainDerived Neurotrophic Factor (BDNF)exerts on both peripheral myelinationby Schwann cells, and central myelinationby oligodendrocytes.Schwann cell myelination: We havemade seminal discoveries showing thatBDNF exerts contrasting influencesupon peripheral myelination. Usingin vitro myelination assays, we havefound that BDNF exerts its influenceupon myelination by acting directlyupon neurons, thereby regulatingaxonal receptivity to Schwann cellmyelination. Despite the fact thatSchwann cells also express receptorsfor BDNF, our data provide no evidencethat BDNF acts directly on Schwwancells to regulate myelination. We findthat the effects of BDNF are dependenton the phenotype of the neuron beingmyelinated, and, in particular, thecomplement of neurotrophin receptorsexpressed by these neurons. We haveutilized a number of highly specialized64

tissue culture techniques, including (i)compartmentalized Campenot chambers,(ii) viral-based shRNA approaches toknock-down neurotrophin receptors, aswell as (ii) mixed genetic co-culturesto investigate the influence that BDNFexerts on myelination in vitro. As a result,we have been able to identify the cellularbasis of how BDNF can both promoteand inhibit the myelination of distinctclasses of peripheral neurons. Thesedata have provided important newinsights into how peripheral myelinationis regulated. Future studies will aim toidentify and dissect the signaling systemsthat BDNF utilizes to regulate thesecontrasting effects.Oligodendrocyte myelination: Wecontinue our investigation into howBDNF regulates the capacity ofoligodendrocytes to myelinate the centralnervous system. Our in vitro data indicatethat BDNF exerts a strong promyelinatinginfluence, and our analyses haveidentified that BDNF directly activatesthe receptor tyrosine kinase TrkBexpressed on oligodendrocytes topromote myelination. We have initiated acollaboration with Professor Luis Parada(University of Texas Southwestern, USA)to access his recently generated TrkBfloxed mice in order to generate micewith an oligodendrocyte-specific deletionof TrkB, and we have commenced ananalysis of the effect this deletion has onmyelin development. The aim of theseexperiments is to verify our in vitro dataand to fully establish the impact that lossof TrkB signaling in oligodendrocytes hason myelin development in vivo.In addition, we have also begun aseries of in vitro experiments aimedat identifying the molecular basis ofthis promyelinating influence. Wehave taken a candidate approach andinterrogated the activation status of anumber of oligodendrocyte cytolosicproteins, such as Erk1, 2, Akt, and thenon-receptor tyrosine kinase Fyn, toinvestigate the impact their activationhas on oligodendrocyte myelination.We will continue this analysis to identifythe key signaling pathways activated byBDNF that promote oligodendrocytemyelination.Collectively, these projects undertake adetailed cellular, molecular and geneticinvestigation into the mechanisms thatcontrol myelination. Such investigationswill provide new insight into, and increaseour understanding of the molecularprocesses that govern the myelinatingprocess. This is a critical step inestablishing how these responses mightultimately be targeted and modulatedfor therapeutic benefit in the context ofdemyelinating disease.Regulation of Neurotrophin SignalingMelanie Willingham, Agnes Wong,Trevor J Kilpatrick and Simon MurrayThe neurotrophins regulate a diverserange of biological effects, and a majortheme of our laboratory’s research is toinvestigate the influence that a numberof related molecules and co-receptorsplay in regulating the signaling of thisimportant growth factor family:(i) The Neurotrophin ReceptorHomolog-2 (NRH2)Our analysis of this novel neurotrophinco-receptor continues. We arecompleting studies that examinethe influence that NRH2 exerts uponneurotrophin signaling and in a cellularmodel of neurotrophin-inducedneuronal differentiation.(ii) pro-Neurotrophin signalingThe precursor forms of theneurotrophins, collectively known asthe pro-neurotrophins, have recentlybeen shown to be biologically active.We have identified a clear role forpro-BDNF in inducing apoptosisin primary Schwann cell cultures,and havd begun to identify thesignal transduction mechanismsutilized. Micro-array data indicatethat proBDNF strongly influencescholesterol metabolism in Schwanncells, and this has been confirmedin subsequent biochemical analyses.We are currently investigatingthe mechanisms of cholesterolregulation, and the role it is playingboth in the context of cell deathand in myelination. These studieshave identified potentially importantnovel functions of pro-neurotrophinsignaling.A follow up to the ANZgene GWAS forMultiple SclerosisTrevor J. Kilpatrick, Helmut Butzkuevenand Judith Field with The Australia andNew Zealand Multiple Sclerosis GeneticsConsortium (ANZgene)The recent genome-wide association scan(GWAS) performed by the Australia andNew Zealand MS Genetics Consortium(ANZgene) led to the identification oftwo new loci associated with the riskof developing multiple sclerosis (MS),and has directed us to further analysisof MS genetic risk, and the functionalconsequences of these genetic changes.The Chromosome 12 locus identifiedas associated with increased risk of MSencompasses 17 genes. Our aim is todetermine which of these genes containsvariation associated with risk of MS, withthe ultimate goal of ascertaining thefunctional consequences of changes tothis gene. This has involved the methodof deep-sequencing of the entire 300kilobase (kb) region of DNA associatedwith MS risk. Efforts have focused onthe amplification of this region using thetechnique of long-range PCR, as wellas pooling of DNA samples from MScases and healthy controls followed byspecific sequence capture methods forthis 300 kb region. These two methodsof DNA enrichment have resulted inDNA that has now been sequenced andis currently being interrogated for novelrare variations that can be analysed forassociation with MS risk.The CD40 locus has previously beenassociated with the risk of autoimmunedisease development, particularly Graves’disease and rheumatoid arthritis. Thevariation associated will all three of theseautoimmune diseases falls within a part ofthe CD40 gene sequence known to affectCD40 protein expression levels, resultingin enhanced CD40 expression in thecase of Graves’ disease and rheumatoidarthritis compared to the MS risk-allele. Inour follow up to the ANZgene GWAS, weare investigating the effects of the CD40risk SNP in MS patients compared tohealthy individuals, focusing on specificimmune cell subtypes found in theblood, with the aim to determine whatthe consequences of these changes inexpression are in relation to the functionof the individual’s immune system.65

Multiple SclerosisFinally, using the ANZgene GWAS datawe investigated the association of singlenucleotide polymorphisms (SNPs) acrossthe Human Leukocyte Antigen (HLA)complex in 1618 MS cases and 3413controls of European ancestry. A total of1927 SNPs within the HLA complex wereinterrogated, with logistic regressionanalysis resulting in the identification of7 SNPs independently associated withMS conditional on the others (p < 4 x 10 -6 ).The association of these 7 SNPs with MSwas independently replicated in a cohort2212 MS cases and 2251 controls(p < 0.001), with a combined p-valueof p < 6 x 10 -8 . The rs9277535 SNP wasfound to be strongly associated withrisk of developing MS in the GWASdiscovery data set (p < 9 x 10 -9 ) and thiswas independently replicated (p < 7 x 10 -4 ;combined p < 2 x 10- 10 ). The rs9277535SNP identifies the HLA class II DPB1 alleleDPB1*0301, which has previously beenassociated with MS in smaller samplesof MS cases and controls. We havetherefore identified an independentMS-susceptibility locus with genomewidesignificance in the HLA class IIDPB1*0301 allele.How do MS risk genes work?Cathy Jensen, Justin Rubio and HelmutButzkueven in collaboration withJ Stankovich, University of Tasmaniaand B Oldfield, Monash UniversityMultiple sclerosis (MS) is a complexautoimmune disease characterisedby demyelinating lesions in thecentral nervous system (CNS). Myelinoligodendrocyte glycoprotein (MOG),a brain-restricted protein expressed onthe surface of myelin, has long beenconsidered a potential target of theabnormal immune response.There are common human geneticvariations in the coding region of theMOG gene that have been associatedwith increased risk of Multiple Sclerosis,in particular the protein substitution520G>A. In order to understand possiblemolecular mechanisms of this association,we have investigated whether expressionor composition (splicing) of MOGencoding RNA is influenced by the riskgenotype in post-mortem human brain.We could not demonstrate any changein expression of MOG RNA driven by therisk genotype. On the other hand, usingquantitative reverse transcriptase PCR(qPCR), we found that the 520A alleleis associated with a significant increasein the way MOG RNA is constructedin the brain, namely in splicing of exon2 to exon 3. This change is likely toincrease levels of cell-surface MOG andreduce soluble MOG. Using predictivealgorithms, we found that the 520G>ASNP alters an exonic splicing enhancer(ESE) sequence involving the SC35 andSRp55 RNA-binding proteins, which couldexplain our results.In conclusion, changes in splicing, butnot expression levels, are associated withcommon genetic variation in the MOGgene. Further work is now required todetermine whether this altered MOGexpression profile, which is predictedto be over-represented in NorthernEuropean MS patients, is relevant to thepathogenesis of this debilitating disease.Do risk genes for MS also changethe rate of disability progression?Cathy Jensen, Trevor J Kilpatrick, JudithField, Justin Rubio, Helmut Butzkuevenand collaboratorsRecent association studies in multiplesclerosis (MS) have identified andreplicated several single nucleotidepolymorphism (SNP) diseasesusceptibility loci including variationsin or near the genes CLEC16A, IL2RA,IL7R, RPL5, CD58, CD40 and a region onchromosome 12q13–14. The last two ofthese were discovered and reported lastyear in the ANZGENE collaboration, withFlorey scientists seminally involved in thediscovery. In this project, we examinedwhether these genetic susceptibilityfactors could also modulate MS diseaseseverity, as demonstrated previouslyfor the MS risk allele HLA-DR15. Weinvestigated this hypothesis in a cohortof 1006 well-characterised MS patientsfrom South-Eastern Australia. Wetested the MS-associated SNPs forassociation with five measures of diseaseseverity incorporating disability, age ofonset, cognition and brain atrophy. Weobserved trends of association betweenthe RPL5 MS risk SNP and time betweenfirst demyelinating event and relapse,and between the CD40 risk SNP andsymbol digit test score, a test of cognitivefunction. No associations were significantafter correction for multiple testing.Therefore, we found no evidence forthe hypothesis that these new MSdisease risk-associated SNPs influencedisease severity, which potentially leadsto the conclusion that there could bedifferent risk factors (both genetic andenvironmental) for the initiation of MSand outcome in MS, once the diseasehas been initiated.StaffTrevor Kilpatrick MB BS PhD FRACPHelmut Butzkueven MB BS PhD FRACPHolly Cate BSc MSc PhDSimon Murray BSc BAppSci PhDToby Merson BA BSc (Hons) PhDJudith Field BSc (Hons) PhD (from July 2009)Michele Binder BSc (Hons) MScMelissa Gresle BSc (Hons) PhDVilija Jokubaitis BCom/BSc (Hons) PhDResearch AssistantsSze Woei Ng MbiotechBioinform BScLaura Johnson BAppSci (from July 2009)Bill Doherty BSc (Hons)Daniel Merlo BSc (Biological Science)Dennis Kemper BSc (Hons)StudentsGerry (Zhi-Ming) Ma BSc (Hons)Jennifer Sabo BA (Bryn Mawr USA) (PhDcandidate)Anna Jonas BSc MSc (Germany) (PhDcandidate)Laura Oluich BBNSc (Hons) (PhD candidate)Cathy Jensen BAppSci (Med Sc) (PhDcandidate from July 2009)AwardsJunhua XiaoSociety for Neuroscience ChapterPost-doctoral Travel Award Societyfor Neuroscience (USA, 2009)66

Conferences and PresentationsTrevor KilpatrickANS 2009, Canberra. Gas6 deficiencyincreases oligodendrocyte loss andmicroglial Activation in response tocuprizone induced demyelination.January 2009 (invited speaker).AusiMed Medical Conference, Israel.Multiple Sclerosis and stem cell therapy.April 2009 (invited speaker).American Academy of Neurologymeeting, Seattle, USA. Genome-WideAssociation Scan Identifies Two NovelMultiple Sclerosis Susceptibility Loci. April2009 (invited speaker).ANZAN Annual Scientific Meeting,Christchurch, NZ. Multiple Sclerosis andstem cells. May 2009 (invited speaker).MSRA progress in MS Conference,Sydney. Genes and the Environment.October, 2009 (session chair).19th World Congress of Neurology,Bangkok, Thailand. Helping to preservepatient functional capabilities in earlyMS: New insights. October 2009 (invitedspeaker and satellite symposium chair).PACTRIMS 2009, Hong Kong. Genes:Australia. November 2009 (invitedspeaker).Helmut ButzkuevenNeuropathology Department, Universityof Muenster, Germany. Glial responses ininflammatory CNS injury: good and bad.September, 2009 (invited speaker).Department of ExperimentalOphthalmology, University of Muenster,Germany. EAE-associated optic neuritisin the mouse: LIF therapeutic effectsScientific Seminar. October, 2009(Invited speaker).University of Bochum, Germany.Neuroscience Seminar: A genomewideassociation study identifies newsusceptibility loci for multiple sclerosis.October, 2009 (invited speaker).PACTRIMS 2009, Hong Kong.MSBase: Powering Australian andInternational Longitudinal Databsingin Multiple Sclerosis. November 2009(invited speaker).Neurology Grand Round, Royal NorthShore Hospital, Sydney. Recent riskgene discoveries in Multiple Sclerosis.November 2009 (invited speaker).Simon MurrayAustralian Neuroscience Society 30thAnnual Meeting, Canberra. ‘Sortiling’it out in neurodegenerative disease:Mechanism and function of receptorendocytosis. January 2009 (co-chair).Vanderbilt University. ‘Wrapping it all up’ –The diverse roles of BDNF in regulatingmyelination. October 2009 (invitedspeaker).Tobias MersonMultiple Sclerosis Research AustraliaProgress in MS Research ScientificConference. Clinical phenotype precedesovert demyelination in a transgenicmodel of inducible oligodendrocyteapoptosis. October 2009, Sydney(Oral presentation).Major Collaborative LinksNationalDr James BourneAustralian Regenerative MedicineInstitute, Monash UniversityNeural precursor cell biologyin the non-human primateAssociate Professor Anne Turnley(Centre for Neuroscience, Universityof Melbourne) and Stefania Castelletto(Melbourne Materials Institute,University of Melbourne)Application of nanoparticle technologyto the study of neural stem cell responsesin vivoCatriona McLeanDirector, Department of AnatomicalPathology, The Royal Prince AlfredHospital, Victoria, AustraliaThe role of Gas6 and the TAMreceptors in central demyelinationDr Rachel Hill and Professor Maartenvan den Buuse, (BehaviouralNeuroscience Laboratory, MentalHealth Research Institute, Victoria)Brain-derived neurotrophic factorexpression is increased in thehippocampus of 5-HT(2C) receptorknockout miceProfessor Gary EganFlorey Neuroscience InstitutesNeuroimaging in MS and in mousemodels of demyelinationAssociate Professor Andrew GundlachFlorey Neuroscience InstitutesRole of Galanin in demyelinationDr Tony HughesDept. Parmacology,University of MelbourneNeurotrophin mimeticsAssociate Professor Graham BarrettDept. Physiology, University of MelbourneRole of the low affinity neurotrophinreceptor in myelinationProfessor Perry Bartlett and ProfessorPankaj Sah (Queensland Brain Institute)Investigation of the functional role ofneural precursor cells in hippocampallearning and memoryProfessor Simon FooteMenzies Research InstituteGenetics of Multiple SclerosisProfessor Tony McMichael (AustralianNational University, Canberra) andAssociate Professor Anne-LouisePonsonby (Murdoch Children’sResearch institute)Role of environmental factors in MS:Ausimmune studyInternationalProfessor Greg LemkeThe Salk Institute for Biological Studies,La Jolla, USAThe role of Gas6 and the TAM receptorsin central demyelination67

Multiple SclerosisProfessor Samuel DavidMcGill University, CanadaIron metabolism in MSProfessor David WynickBristol University, UKRole of Galanin in demyelinationAssociate Professor Bruce CarterVanderbilt University, USANeurotrophin signaling inmyelinating cellsProfessor Luis ParadaUniversity of Texas Southwestern, USARole of TrkB in central demyelinationProfessor Gerry ShawMcKnight Brain Institute of theUniversity of FloridaBiomarkers of neuronal damageand disease severityAssociate Professor Tanja KuhlmannInstitute of Neuropathology, UniversityHospital Münster, Münster, GermanyRole of Dab2 in central demyelination68

Neurodegeneration— Molecular NeuropharmacologyResearch OverviewThe research endeavours of the MolecularNeuropharmacology team addresscellular mechanisms underlying injuryprocesses that contribute to acute andchronic neurodegenerative conditions.Whilst for many years a “neuro-centric”view of neurotransmission and braindysfunction was paramount, glia arenow considered to play major roles inbrain physiology and pathology. Ourrecent focus has not only been on bothneurones and astrocytes, the mostpopulous cells in brain, but also ontheir interaction and how excitotoxicity,oxidative stress and inflammationas generic mechanisms consideredcausative in neurodegenerationaffect these two cellular populations.Excitotoxicity is a destructive processinvolving neuronal death by apoptosisand necrosis, subsequent to the overstimulationof receptors for the brain’smajor excitatory transmitter, L-glutamate(Glu). Rises in intracellular calciumactivate an amplifying cascade of toxicevents involving free radical generationand mitochondrial dysfunction inneurones. Astrocytes play integral rolesin the maintenance of brain functionand synaptic transmission throughtheir involvement in energetics andmetabolism, secretion of trophic factorsand transport of various molecularspecies. Inflammation is another majorcomponent of astrocyte biology andcan be both protective or destructive,dependent upon the strength of theinjurious response.We employ a wide range of strategiesand experimental approaches to studythe integrated involvement of neuronesand astrocytes in brain injury. Continuingmajor contributions have been madeto the understanding “mitochondrialdeath” signalling associated with braindysfunction. Detailed analyses of theredistribution of apoptogenic proteins indifferent populations of neurones at risk inhypoxic ischaemic injury, Parkinson’s andHuntington’s diseases, have allowed usto understand how the temporal patternsof activation of various destructiveproteases can induce downstreamelimination of neurones. Importantly, wefound that caspase-independent celldeath involving calpains is extremelyimportant when injury is elicited bypathological stressors. Our work hasdefined a contemporary view of neuronalinjury wherein programmed cell death(PCD) occurs with differential, incompleteactivation of the intrinsic mitochondrialpathway to allow patterns of injury thathave varying mixes of downstreamactivation of caspase-dependent and-independent mechanisms. We now viewPCD as an interactive mix of the intrinsicmitochondrial cascade, autophagy andprogrammed necrosis, and which arevariably recruited dependent upon insulttype and neuronal phenotype.The neurobiology of astrocytes isundergoing a remarkable renaissancesuch that this largest population ofcells continues to surprise by displayingan incredible spectrum of biologicalresponses. Astrocytes outnumberneurones in mammals and displayvarious forms of plasticity – many ofthese responses target maintenanceof neuronal life and rescue threatenedneurones. Various efforts in our teamhave provided insights into thepatterns of involvement of astrocytesin the potentially destructive cellularmilieu. Firstly, astrocytes can be “preconditioned”to switch on protectivemechanisms downstream of hypoxiainducibletranscription factor (HIF-1).Pre-conditioning stimuli that activateHIF synthesis in astrocytes, stimulatea cascade of protective responses,including one regulating vascularendothelial growth factor (VEGF). ThisHIF-1/VEGF response is one featureof a spectrum of astrocytic responsesthat are cytoprotective as part of theearly phase of astrogliosis. Secondly,astrogliosis also leads to cytoskeletalchanges involving alterations inmorphological phenotype and thesestructural changes appear to involveredistribution of glutamate transporters(EAATs) between intracellular pools andthe cell surface. Using a multidisciplinaryapproach we found maintenance offunctional EAAT proteins at the astrocyticsurface is a further neuroprotectivemechanism (removing toxic Glu fromsynapse), and is a homeostatic processthat is neuroprotective in the face oftoxic events that occur with advancing“chronic” astrogliosis. Thirdly, we believethat if astrocytes are not maintained ina “healthy” phenotype that exposure todiverse toxic stimuli (eg energy deficits,free radicals, protein aggregates) inducesthem to make major contributionsto the pathological mechanisms ofneurodegenerative conditions. Indeed,we have evidence that such astrocyticresponses contribute to the death ofmotoneurones in MND, but also that thepro-survival responses of astrocytes canbe harnessed to switch on endogenoushomeostatic processes that minimiseneurodegeneration.Major Research ProjectsNeuropathology of motor neuronedisease: astrocytes are the nonneuronalneighbours contributing tomotoneurone injuryRoss O’Shea, Nicole Wallis, ChrissandraZagami, Julie Atkin, Manal Farg,Malcolm Horne and Philip BeartMotor neurone disease (MND) isa fatal degenerative condition forwhich there is no effective remedy.New, successful therapies need tobetter target pathological processesin MND. Excitotoxicity contributesto motoneurone injury in MNDand glutamatergic transmission iscompromised in ALS, with in particularthe key astrocyte-based glutamatetransporter (EAAT2) being reduced in apathologically related manner. Elevationof EAAT2 levels delays the death ofmotoneurones suggesting a key role forthe transporter and excitotoxicity in thepathology of MND.For a number of years we have probedthe role of EAATs in motor neurone injury.Since motoneurones are also remarkablysensitive to excitotoxic injury mediatedvia the AMPA subtype of Glu receptor, weexplored the patterns of injury inducedby excitotoxic and oxidative insults inprimary cultures of mouse spinal cordcontaining both motoneurones andastrocytes. In both types of injury EAATtransport was conserved initially andfunctioned in a homeostatic manner tominimize neuronal injury. Differentialprofiles of injury produced by oxidativeand excitotoxic insults identified twodistinct phases of injury which parallelimportant aspects of the pathology ofMND – excitotoxic and oxidative insultspreferentially affected motoneuronesand astrocytes, respectively. Combinedoxidative and excitotoxic insultsproduced time- and concentrationdependentmorphological changes that70

paralleled reductions in cellular viability,and effected a more rapid inductionand a greater extent of injury than eitherinsult alone. EAAT2 immunoreactivityassociated with astrocytic processeswas dramatically increased at 24 hr butby 48 hr had decreased below controllevels. However, data strongly implicate ahomeostatic role of EAAT2 in maintainingthe health of the selectively vulnerablemotoneurones – EAAT2 and GFAPimmunoreactivity were maintained wellbeyond that seen for the motoneuronemarker SMI32. The blebbing of neuriticprocesses and the overall slow diebackof motoneurone arbour were stronglyindicative of a substantial involvement ofapoptosis. Astrocytes adjacent to injuredmotoneurones showed heavily reducedGFAP immunoreactivity and underwentastrogliosis. These findings highlightthe importance of the motoneuroneastrocyteinterdependency – lossof astrocytes adjacent to motoneuronesmay compromise their healthpromoting axonopathy.Our findings focus attention on astrocytesas major players in the course of thepathological process. Indeed, astrogliosiswith hypertrophy is well documentedearly in the pathology of both human ALSand rodent models of the disease. Sincepotentially toxic aggregates of mutantCu/Zn-superoxide dismutase (mSOD1)may be causative in both familial andsporadic MND, we hypothesized thataggregation of mSOD1 in astrocytesmight contribute to the pathologicalprocess. This idea is consistent with“non-cell autonomous” concept ofmotoneurone injury, given the ubiquitousexpression of SOD1 in nervous tissue, andlikely effects of SOD1 in neighbouringnon-neuronal cells viz. astrocytes. Wehave now demonstrated convincingly thatmSOD1 is able to aggregate in astrocytesin a species- and time-dependent manner.Although mSOD1 inclusions wereinitially classified as “micro” = sparselydistributed, or “macro” = concentrated,detailed morphological analyses revealeda much more diverse range of inclusionsrelated to the “clumpiness” and areaof cell occupied. Eventually, a rankingsystem was deemed the most accuratemethod for analysis, and fluorescentmicroscopy allowed the definition ofinclusions as diffuse or mostly diffuse(24 hr), but by 72 hr, there were veryfew diffusely labelled astrocytes, andinstead the majority of the inclusionswere larger, “clumpier” inclusions thatoccupied over 25% of the apparent cellarea. In a parallel fashion image analysisand ranks of morphologies showedthat astrocytes could be categorisedas being “resting”, in the beginningstages of becoming “reactive”, or inthe final stages of taking on a “reactive”phenotype. A most interesting findingwas that cells in the vicinity of mSOD1-EGFP transfected astrocytes also overtime altered their morphologies sothat by the time of maximum mSOD1aggregation this population possesseda distinctly different distribution existingin a phenotype skewed towards the“reactive”phenotype. Indeed, there isevidence that astrocytes can releasenumerous neurochemicals (Glu, nitricoxide, chromagranin A, NGF, Fas-ligand),which are likely to affect the viability ofmotor neurons. Our investigations offerunique insights into the neuropathologyof MND and may provide new clues toits management.Roles and regulation of glutamatetransporters in the central nervoussystemRoss O’Shea, Linda Lau, Rebecca Sheean,Victoria Perreau, Holly Cate, and PhilipBeart in collaboration with Steve Cheungand Jessica Chen (Menzies Institute,University of Tasmania), and LorraineBischof (CSIRO Mathematical& Information Sciences)Glutamate transporters (EAAT1-5)regulate the extracellular concentrationof Glu, the major excitatory transmitterof mammalian brain. Astrocytic EAATS(EAAT1/2) are functionally dominantplaying key roles not only in excitatorytransmission, but additionally in Glu/glutamine metabolism and energetics,and thus represent an essential partof the astrocytic biological repertoire.Dysfunction of Glu transport meansextracellular levels of Glu rise inducingexcitotoxic injury, which is important inischaemia and MND. Altered expressionor function of EAATs has been identified inmany neurological conditions. Astrocytesare very plastic cells, possessing capacitiesto change their volume, and to contractand distend their processes, includingtowards active synapses and in responseto the inflammatory milieu arising inneuropathologies. We have providedevidence for an interdependencybetween EAAT function and the astrocytephenotype and cytoskeleton involvingalterations in transporter activity and/or localization. Mechanisms regulatingthe abundance and function of astrocyticEAATs are poorly understood, butdirectly relevant to understanding howEAATs might be regulated to providebenefit in neurological conditionswhere their function is compromised.Primary astrocytes in culture offer greatadvantages to study such interfacesand we have used a multi-facetedstrategy employing pharmacological andmolecular approaches, in concert withimmunocytochemistry and advancedimage analysis, to probe the regulationof EAAT function.We have found that the astrocytedisplays a homeostatic mechanismserving to normalize and/or maintainEAAT function. In particular, the cellsurface expression of EAAT2 is conservedlikely because it is an astrocytic proteinpresent at remarkably high abundance(~1% of proteome), and intimately linkedto Glu metabolism and bioenergetics.Our morphological observationsdemonstrated prominent changes in thepatterns of immunolabelling for the keyastrocytic marker GFAP and EAAT2. Wefind treatment-dependent morphologicalchanges across a continuum ofphenotypes wherein cobblestone andstellate morphologies were at oppositeends of this distribution. These changespresumably reflect differential recruitmentof signalling cascades directly or indirectlyregulating the cytoskeleton, and the neteffect likely involves actin re-organization,especially as we see frequent changesin labelling for F-actin. In astrocytes theactin cytoskeleton is pivotal to manyfunctions and Rho GTPases (Rho, Rac,Cdc42) are linked to actin-rich structuressuch as lamellipodia and filopodia,fundamental determinants of motility andmigratory potential. Indeed, treatment ofastrocytes with the Rho kinase inhibitor,Fasudil (HA-1077) produced rapidstellation in 30-60 min and slow elevationover 24-72 hr of Glu uptake with a 2-foldincrease in the cell surface expression ofEAAT2. Morphological analyses using anewly implemented, automated strategyrevealed alterations to both astrocyticterritories and structure. When geneprofiles were subjected to bioinformaticanalyses, substantial changes were foundin the biological processes associatedwith the actin cytoskeleton, axonguidance, TGFγ-signalling and tight71

Neurodegeneration— Molecular Neuropharmacologyjunctions. Many of genes up- and downregulatedhave cytoskeletal roles andcontribute to the morphological changesinduced by Fasudil . Target validationof a number of genes by quantitativeRT-PCR confirmed the changes found bymicroarray, including the up-regulation ofEAAT2. Particularly incisive was evidencethat Fasudil-induced stellation induceda “prosurvival” phenotype involvingnot only EAAT2 up-regulation, but alsoelevated expression of anti-oxidant,trophic factor and glycolytic genes.Within these transcriptomic data therewere expression changes in a number ofgenes likely to involved in the assemblyof an EAAT molecular complex, includingsubunits of the Na,K-ATPase, Nherf1,Nhe1 and endothelin receptor-B. Theseproteins, and other potential componentsof a cytosolic scaffold such as glutathione,glutamine synthetase and glycolyticenzymes, will be the subject of futurestudies. While studying the actions ofrotterlin, a broad spectrum inhibitorof protein kinase C (PKC), we found arapid decrease in EAAT function, but asurprising increase in the cells surfaceexpression of EAAT2 consistent with thehomeostatic mechanism described above.Endosomes, endoplasmic reticulum,Golgi apparatus and proteosomesappeared not to contribute to themaintenance of EAATs at the cell surface.Surprisingly, monensin an activator ofNa + -ionophore cellular influx acceleratedthe recovery from the inhibition of Gluuptake induced by rotterlin. This findingfocused out attention on Na,K-ATPasewhich sparse reports have linked to Glutransport. Using a newly establishedprocedure reflecting the activity of theNa,K-ATPase we found that rotterlininhibited ouabain-sensitive 86 Rb uptake.Reduced Na + entry would causeinhibition of EAAT activity, which is welldocumented to be Na + -dependent.Monensin returned Glu uptake to controllevels by stimulating Na+ entry as shownby increased 86Rb uptake. How Na,K-ATPase regulates EAAT activity remainsto be resolved, but its α1 subunit ispresent in astrocytes and we found bothits ATPB-1 and FXYD5 components downregulatedin our microarray work. Theselinkages and any partial-decoupling ofEAAT-Na,K-ATPase may underlie thecontinuum of homeostatic responses wehave observed in EAAT function and arethe subject of ongoing work.Mitochondrial contributions to diversemodes of neuronal injuryPhilip Beart, Linda Mercer, Ross O’Shea,Percy Chu, Linda Lau, and Yea Seul Shinin collaboration with Phillip Nagley,Gavin Higgins and Rod Devenish(Monash University)Neurones undergo injury and eventualdeath in a variety of ways during braindysfunction and mitochondria play a keyrole in such outcomes. In pathologicalsettings such as hypoxic-ischaemic injury,Parkinson’s and motoneurone diseases,mitochondria in affected cells, beingcentral to cellular homeostatic andenergetic activities, play key regulatoryroles in death processes. Variousdestructive mechanisms, includingenergy deficits, toxic free radicals andexcitotoxicity, converge on mitochondria,which are considered “guardians atthe gate of injury”. Recent advances inour understanding of neuronal injuryindicate that permeabilisation of theouter mitochondrial membrane andredistribution of apoptogenic proteins(cytochrome c, Smac, AIF, HtrA2, EndoG) enable progression of programmedcell death (PCD). However, classicalcaspase-dependent apoptosis (PCDtypeI) is now recognized to be a lesscommon event than initially believed inneuronal injury. Instead there is sharing of“death” machinery and crosstalk betweensignalling wherein neuronal injury caninvolve components of apoptosis,caspase-independent programmednecrosis (PCD-type III) and autophagy(PCD-type II). While post-mortem humanbrain tissue and animal models of diseaseprovide clues to the diversity of neuronaldeath, the clearest insights into molecularmechanisms come from laboratory studiesusing cellular models of injury or disease.In a long-term programme we haveexamined how the neuronal phenotype,type of stressor and intensity of stressdetermine the recruitment of deathsignalling and the nature of eventualdemise in primary cultures of neurones.Energy deficits, toxic free radicals andexcitotoxicity can all disturb mitochondrialfunction activating death signalling. Thuswe found that pharmacological inhibitionof the respiratory chain using rotenone,3-nitroproprionic acid, antimycin A andpotassium cyanide, which are respectivelyinhibitors of complexes I, II, II and IV,produced a slow pattern of neuronalPCD across the apoptotic-necroticcontinuum. Regardless of the type or mixof PCD, injury is hierarchical and exhibitsprecise temporal dependency. Thus forexample trophic-factor deprivation incerebellar granule cells produces classicalPCD-type I with caspase-3 activationand early release of cytochrome cand Smac, but the oxidative stressorhydrogen peroxide produces a mixedprofile of injury involving both caspaseand calpain activation but earlier releaseof the caspase-independent “death”proteins AIF and HtrA2. In neurones,cross-talk between caspase-dependentand caspase-independent pro-apoptoticsignalling seems the norm rather than theexception. Striatal GABAergic neuronesexhibit canonical, insult-dependent PCDwith early redistribution of caspaseindependent“death” proteins whenchallenged with a wide variety of stressors.In general, mobilization of Bax is a keytrigger of the mitochondrial recruitmentand “death” signalling, and changes inmitochondrial transmembrane potentialare less common after exposure todiverse insults.Recent work has dissected aspects ofneuronal PCD produced by varyingdegrees of oxidative stress since thisprocess is well recognized to contributeto neuropathologies. Further analysesof neuronal injury induced by hydrogenperoxide confirmed a type of PCD quitedistinct from apoptosis and revealed theinvolvement of programmed necrosis(PCD-type III). Injury was more rapid thanapoptosis, caspase- and time-dependent,and involved loss of plasma membraneintegrity and DNA fragmentation.Silencing of Endo G, but not AIF, largelysuppressed cell death. We also founda component of autophagy that couldbe attenuated by treatments directedat this type of PCD-type II - 3-methyladenineand silencing of the autophagygene Atg7both reduced injury. Theappearance of autophagosomes was alsolargely attenuated by 3-methyl-adenine.In parallel work examining apoptoticinjury, there was differential involvementof autophagy likely to be linked to theremoval of damaged proteins and notcell death. Longer term exposure to morechronic oxidative stress engendered aprofile of injury without caspase activation,in which there was a more rapidmembrane permeabilization and DNAfragmentation. Here 3-methyl-adenine72

was ineffective and silencing Atg7 onlytemporarily delayed cell death, as didsilencing of Endo G. Thus in chronicoxidative stress autophagy (PCD-typeII) and programmed necrosis (PCD-typeIII) were overwhelmed by unregulatednecrosis – although interestingly celldeath events were still ordered.Moreover, very similar patterns of injurywith parallel recruitment of differenttypes of PCD have been described inanimal model of focal brain ischaemia.Thus our findings have implications forcerebral hypoxic-ischaemic injury afterstroke where if oxidative stress can beameliorated by drug regimens thenthe resultant extent of brain damagecould be minimized. Our currentbelief is that there is insult-dependentsharing of death pathways whereinactivation of the intrinsic mitochondrialcascade, autophagy and programmednecrosis represent a destructive “triad”,variably recruited dependent uponcellular status, extracellular milieu andneuronal phenotype.Other Research ProjectTolerance in the brain: studyingendogenous mechanisms which canminimise brain cell death in models ofperinatal asphyxiaPhilip Beart, Percy Chu, and Linda Mercer,in collaboration with Nicole Jones(University of NSW)Hypoxia is major contributor to theresultant brain damage in neurologicaland neurodegenerative conditions,including stroke and perinatal asphyxia.Various clinical reports document thatpre-treatment (or preconditioning) witha mild, non-damaging exposure tohypoxia can protect against a subsequentbrain injury. Many researchers believethat the transcription factor, HIF-1, is akey regulator of adaptive responses tohypoxia since it regulates expressionof target genes such as erythropoietin,VEGF, glycolytic enzymes and glucosetransporters. These proteins haveneuroprotective actions and are involvedin mediating adaptive processes suchas erythropoiesis, angiogenesis andincreased glucose transport to the brain.HIF-1 activity has recently been shownto be modulated by a group of prolylhydroxylase (PHD) enzymes. There areseveral drugs (PHD inhibitors) that canstimulate the production of the HIF-1protein, several HIF-1 target genes andthe downstream adaptive pathwaysdescribed above. We adopted an uniqueapproach by selectively targeting HIF-1/PHD function to repair the brain. Weinvestigated functional and pathologicaloutcomes after hypoxic-ischemic (HI)brain injury in the infant rat. Behaviouraldeficits are found after HI and thesedeficits are accompanied by an unilaterallesion compared to sham control animals.Hypoxic preconditioning and the PHDinhibitors desferrioxamine (DFX) andcobalt chloride, 24 hr prior to HI, reducedneuropathological and behaviouraldeficits associated with HI injury 5 weeksafter HI. In other studies, we definedmechanistic aspects of preconditioninginvolving HIF-1 in vitro employingvarious cell culture models and studiedprotective actions of preconditioningwith PHD inhibitors in cortical astrocytes.We demonstrated that preconditioningwith the PHD inhibitors DFX and ethyl-3,4-dihydroxybenzoate protectedastrocytes against oxidative stress viaa HIF-1/VEGF-dependent mechanism.Astrocytic HIF-1α expression and VEGFrelease thus influence the function ofsurrounding cells and vasculature duringneurological insults. These insights into inendogenous neuroprotection of the brainreveal new targets for the design of noveltherapeutic strategies for treating strokeand perinatal asphyxia.Staff ListFellows, Senior Research Officers andResearch OfficersPhilip Beart BSc (Hons) (Adel) PhD (ANU)DSc (Melb)Ross O’Shea BSc (Hons) PhD (Melb)Linda Mercer BSc (Hons) (ANU) MSc (Mon)Linda Lau BBiomedSc (Hons) (Monash) PhD (Melb)Research AssistantsPercy Wai Chu BBiomed Sci (Hons) (Mon)(until April 2009)StudentsPercy Wai Chu BBiomed Sci (Hons) (Mon)Nicole Wallis BSc (Hons) (Melb)Rebecca Sheean BSc (Hons) (La Trobe)Yea Seul Shin BSc (Hons) (Auckland)Conferences andPresentationsPhilip BeartAustralian Neuroscience Society,Canberra, 2009American Society for Neurochemistry,Charleston, 200911th International Neuroscience WinterConference, Sölden, Austria, 2009European Society for Neurochemistry,Leipzig, Germany, 2009International Society for Neurochemistry,Busan, Korea, 20099th ISN Advanced School ofNeurochemistry, GyoengJu, Korea, 2009European Cell Death Organization,Paris, 2009Australasian Society for Clinical andExperimental Pharmacology andTherapeutics, Sydney, 2009Ross O’SheaAustralian Neuroscience Society,Canberra, 2009International Society for Neurochemistry,Busan, Korea, 2009Linda MercerIBRO Congress, Melbourne, 2007Australian Neuroscience Society, Hobart,2008Asia-Pacific Society for Neurochemistry,Shanghai, 20083rd ISN Special NeurochemistryConference, Beijing, 2008Linda LauAustralian Neuroscience Society, Hobart,20089th ISN Advanced School ofNeurochemistry, GyoengJu, Korea, 2009International Society for Neurochemistry,Busan, Korea, 200973

Neurodegeneration— Molecular NeuropharmacologyMajor Collaborative LinksNationalProf Phillip NagleyDepartment of Biochemistry,Monash University, MelbourneMitochondrial contributions toneuronal injuryProf Rod DevenishDepartment of Biochemistry,Monash University, MelbourneAutophagy and mitochondriain neuronal injuryDr Nicole JonesDepartment of Pharmacology,University of NSWPreconditioning the brainand cytoprotectionProf David PowDepartment of Anatomy,University of QueenslandCellular localisation ofglutamate transportersDr Steve CheungMenzies Research Institute,University of TasmaniaTranscriptomic analyses ofgenes in neuronal injuryProf Sheila CrewtherSchool of Psychological Sciences,La Trobe UniversityRetinal oxygen and hypoxicinducible factorInternationalProf Arne SchousboeDanish University of PharmaceuticalSciences, DenmarkAnalyses of the roles of glutamatetransportersProf Niels DanboltDepartment of Anatomy,University of Oslo, NorwayCellular localisation of glutamatetransportersDr Thelma LovickDepartment of Physiology,University of Birmingham, UKCholecystokinin receptors in painand analgesiaDr Marcus RattrayUniversity of Reading, Reading, UKMolecular neurobiology of glutamatetransportersEditorial PositionsPhilip BeartBritish Journal of PharmacologyPublicationsBeart PM, Balcar VJ. (2009) GrahamJohnston: bringing success toneuroscience through medicinalchemistry. Neurochem Res 34:1696-1697.Diwakarla S, Nagley P, Hughes MLR, ChenB, Beart PM (2009) Differential insultdependentrecruitment of the intrinsicmitochondrial pathway during neuronalprogrammed cell death. Cell Mol Life Sci66: 156-172.Diwakarla S, Mercer LD, Kardashsyan L,Chu PWY, Shin YS, Lau CL, Hughes MLR,Nagley P, Beart PM (2009) GABAergicstriatal neurons exhibit caspaseindependent,mitochondrially mediatedprogrammed cell death. J Neurochem109 (Suppl 1): 198-206.Farso MC, O’Shea RD, Beart PM(2009) Evidence group I mGluR Drugsmodulate the activation profile oflipopolysaccharide-exposed microgliain culture. Neurochem Res (2009) 34:1721–1728.Higgins GC, Beart PM, Nagley P (2009)Oxidative stress triggers caspaseindependentdeath: endonuclease Ginvolvement in programmed cell deathtypeIII. Cell Mol Life Sci 66: 2773-2787.Moldrich RX, Dauphinot L, Laffaire J,Vitalis T, Yann H, Beart PM, Rossier J,Vivien J., Antonarakis SE, Gehrig C, LyleR, Potier M-C. (2009) Proliferation deficitsand gene expression dysregulationin Down syndrome (Ts1Cje) neuralprogenitor cells cultured fromneurospheres. J Neurosci Res 87:3143-3152.Zagami CJ, Beart PM, Wallis N, NagleyP, O’Shea RD (2009) Oxidative andexcitotoxic insults exert differential effectson spinal motoneurons and astrocyticglutamate transporters: Implications forthe role of astrogliosis in amyotrophiclateral sclerosis. Glia 57: 119-135.74

Neurodegeneration— Neural Development and RegenerationResearch OverviewThe Neural Development andRegeneration laboratory, headed byDr Clare Parish and Dr Lachlan Thompson,has a abroad research interest relatingto repairing the injured brain. Thereare 5 major research themes runningwithin the laboratory: understandingdevelopment of dopamine neurons;improving stem cell therapies for neuralinjuries; restoring circuitry in the injuredbrain; exploiting biomaterials to promoteneural repair; and promoting forebrainneurogenesis for neural repair.Major Research ProjectsUnderstanding axon guidancein dopamine developmentAn ongoing challenge for biologistsis to understand the intricate andprecise wiring achieved during braindevelopment. The same accurate wiringis also required to repair the injured brain.We have observed changes in dopamine(DA) plasticity in many pathologies in theadult brain including Parkinson’s disease(PD), addiction and following treatmentwith antipsychotic drugs. The problems ofDA axon guidance are exemplified by theattempts at repairing the degeneratingpathways in PD through cell replacementtherapy. Success of cell replacementtherapy will depend on whether new DAcells integrate with the host circuitry andmake appropriate connections.Unfortunately, the molecular basis ofdopamine (DA) axon guidance duringdevelopment and repair of the diseasedbrain remains poorly understood. Weare investigating new proteins in thiscontext, with a particular focus on Wntsand their receptors. We have shownthat several Wnts have a temporaland spatial expression that correlateswith development of the DA pathways.Furthermore, Wnts can promote DA axongrowth in vitro and in vivo and providedirectional cues for developing axons. Weare confirming many of these findings invarious Wnt related transgenic mice aswell as currently investigating the abilityof Wnts to promote axon growth fortransplanted DA neurons in PD mice. Anunderstanding of the signalling in DAaxon growth and guidance will markedlyincrease our understanding of DAneuronal connectivity. Such knowledgeis not only important for developmentalbiology, but may provide new insightsinto a number of dopamine relateddiseases and present new avenues toimprove cell replacement therapy forParkinson’s disease.Stem cell therapies for DiseaseCell-based repair of the damaged brainremains one of the most highly anticipatedapplications for stem cells in medicalscience. Remarkable progress has beenmade in the stem cell field over thelast decade and there are now routineprocedures for generating a diverse rangeof neuronal phenotypes from pluripotentstem cells in vitro. A major challengeremains in translating this success to an invivo setting, whereby specific populationsof neuronal progenitors generated fromstem cells can be safely and effectivelytransplanted to the injured brain in orderto replace damaged circuitry and restorebrain function. Our laboratory is engagedin an ambitious but focused researchprogram aimed at repairing neural circuitryin animal models relevant to a number ofhuman conditions based upon our abilityto generate three major populations ofprojection neurons from stem cells, andto model human conditions where thereis a need to replace these cell populations,including: midbrain dopamine neuronsthat degenerate in Parkinson’s disease(PD); cortico-spinal projection neuronsdamaged through traumatic brain injuryand stroke; and striatal medium spinalprojection neurons that are damagedin stroke and Huntington’s disease (HD).One of our approaches to improve stemcell therapies includes a cell sortingstrategy to isolate only desired cell-typesfrom a heterogeneous population for thetreatment of PD. This approach enablesthe elimination of potentially tumorogeniccells or cells that cause unwanted sideeffects. In collaboration with researchersat the University of Melbourne (MarkDenham and Mirella Dottori) we arealso examining the remarkable ability ofhuman embryonic stem cells to integrateand reconstruct long distance pathwaysin the host brain. We have preliminaryfindings illustrating that these cells candelay disease onset in a rat model ofmotor neuron disease and could have thepotential to improve function in rodentmodels of traumatic brain injury and stroke.Biomaterials to support neuronsand promote brain repairIn collaboration with the MaterialsEngineers at Monash University (JohnForsythe and David Nisbet). Repair ofthe diseased or injured central nervoussystem will depend on the capacity todeploy replacement cells and providingappropriate signals for these new cells todifferentiate and integrate and into thehost circuitry. Promoting the survival andneuronal connectivity of new cells willrely on establishing appropriate chemicaland physical environments to supportneural cells and their processes. This willinclude structural elements reminiscentof the extracellular matrix, trophicproteins and suppression of inhibitorymolecules. We are investigating theability of 3-dimentional biomaterials toprovide physical and chemical support forneural stem cells in vitro. We have shownthat modified scaffolds can improve thesurvival, proliferation and differentiation ofstem cells as well as support axon growth.We are able to immobilize proteins ontothese scaffolds to enhance cell support.We are currently investigating theability of these biomaterials to supporttransplanted stem cells in the brain andrestore neural circuitry.Other Research ProjectsRestoring the nigrostriatal pathwayin Parkinson’s DiseaseManipulation of the neurogeniczones to facilitate self-repair inthe Huntington’s disease orstroke-damaged striatumStaff ListFellows, Senior Research Officers,and Research OfficersClare Parish BBiomedSci (Hons), PhD (Monash),NHMRC CDA FellowLachlan Thompson BSc (Hons), PhD (Monash)Chris Bye BSc (Hons) (UNSW)Chathurini Fernando BSc (Hons), MSc(Auckland) PhD (Melb)Research and Technical AssistantsDoris Tomas BSc (Deakin)Mong Tien BSc75

Neurodegeneration— Neural Development and RegenerationStudentsBrette Blakely BA (Hons) (Wellesley, USA),MA (Monash)Suresh Kaushik BSc (Hons)Jerani Pettikiriarachchi BSc BEng (Hons)(Monash)Conferences andPresentationsClare Parish19th Annual Meeting of Network ofEuropean CNS Transplantation andRestoration (NECTAR) Cardiff, WalesAustralian Neuroscience Society 29thAnnual Meeting, Canberra, AustraliaNeurosciences Victoria seminar seriesLachlan Thompson19th Annual Meeting of Network ofEuropean CNS Transplantation andRestoration (NECTAR) Cardiff, WalesChris ByeAustralian Neuroscience Society 29thAnnual Meeting, Canberra, AustraliaMajor Collaborative LinksNationalDr John Forsythe and Dr David NisbetDepartment of Chemical Engineering,Monash UniversityBiomaterials and scaffoldsProf Collin PoutonMonash UniversityCell based therapies for Parkinson’sDiseaseDr Mirella Dottori/ Dr Mark DenhamThe University of MelbourneRepairing neural circuitry using humanembryonic stem cellsProf Stephen StackerLudwig Institute for Cancer Research,Parkville, AustraliaWnt/Ryk in dopamine developmentInternationalProf Anders BjorklundDept. of Experimental Medical Science,Lund UniversityCell Therapy for Parkinson’s diseaseProf Deniz KirikDept. of Experimental Medical Science,Lund UniversityCell and Gene Therapy for Brain RepairDr Jesper JorgensenNsGene Inc., Copenhagen, DenmarkNovel Trophic Factors for Brain RepairProf Evan SnyderBurnham Institute forMedical Research, USACell based therapies for Parkinson’sDiseaseProf Ernest ArenasDept Medical Biochemistry andBiophysics, Karolinska Institute, SwedenWnts signalling in dopaminedevelopmentAssoc Prof Jan Stenman,Ludwig Institute of Cancer Research,Stockholm, SwedenWnt signalling in DA developmentVisiting ScientistsProf Deniz Kirik MD, PhDLund University, SwedenDr Asa Peterson MD, PhDLund University, SwedenAwardsClare ParishAustralian Genome Research FacilityBioplatforms AwardBrette BlakelyAyse Berke & Liana Colvill Travel AwardPublicationsC.L. Parish*, E. Lonardo*, S. Ponticelli,D. Marasco, D. Ribeiro, M. Ruvo,S. De Falco, E. Arenas, G. Minchiotti.A synthetic cripto blocking peptideimproves embryonic stem cellreplacement therapy in Parkinsonian rats.Stem Cells Accepted 27 May (2010).Grealish S, Jönsson ME, Li M, Kirik D,Björklund A, Thompson LH. The A9dopamine neuron component in graftsof ventral mesencephalon is an importantdeterminant for recovery of motorfunction in a rat model of Parkinson’sdisease. Brain. 2010 Feb;133 (Pt 2):482-95.C.V Fernando, D. Moses., C. Parish, D.Tomas, J. Drago, M.K. Horne. Creatinga ventral midbrain stem cell niche inthe Parkinsonian brain. Stem Cells andDevelopment In Press 23 Apr (2010).Pettikiriarachchi, J.; Parish, C.L.; Nisbet,D.R.; Forsythe, J.S. Morphologicaland Chemical Functionalisation ofNanofibrous Scaffolds for TissueEngineering. Tissue Engineering ReviewIn Press (2010)Jerani T. S. Pettikiriarachchi, ClareL. Parish, Molly S. Shoichet, John S.Forsythe, David R. Nisbet. Biomaterialsfor brain tissue engineering. AustralianJournal of Chemistry, In Press (2010).Walker AK, Farg MA, Bye CR et al. Proteindisulphide isomerase protects againstprotein aggregation and is S-nitrosylatedin amyotrophic lateral sclerosis.Brain.133:105-116. (2010)Thompson LH, Grealish S, Kirik D,Björklund A. Reconstruction of thenigrostriatal dopamine pathway in theadult mouse brain. Eur J Neurosci. 2009Aug;30(4):625-38.Thompson LH, Björklund A. Transgenicreporter mice as tools for studiesof transplantability and connectivityof dopamine neuron precursors infetal tissue grafts. Prog Brain Res.(2009);175:53-79. Review.Jönsson ME, Ono Y, BjörklundA, Thompson LH. Identification oftransplantable dopamine neuronprecursors at different stages ofmidbrain neurogenesis. Exp Neurol.2009 Sep;219(1):341-54.76

Friling S, Andersson E, Thompson LH,Jönsson ME, Hebsgaard JB, Nanou E,Alekseenko Z, Marklund U, KjellanderS, Volakakis N, Hovatta O, El ManiraA, Björklund A, Perlmann T, Ericson J.Efficient production of mesencephalicdopamine neurons by Lmx1a expressionin embryonic stem cells. Proc Natl AcadSci USA. 2009 May 5;106(18):7613-8.M.K. Horne, D.R. Nisbet, J.S. Forsythe,C. Parish. Three dimensional nanofibrousscaffolds incorporating immobilized BDNFpromote proliferation and differentiationof cortical neural stem cells. Stem Cells &Development. In Press 15 Oct (2009).L. Cajánek, D. Ribeiro, I. Liste, C.L.Parish, V. Bryja, E. Arenas. Wnt/ßcateninsignaling blockade promotesneuronal induction and dopaminergicdifferentiation in embryonic stem cells.Stem Cells. 27(12):2917-27. (2009).P. Sacchetti, K.M. Sousa, A.C. Hall, I. Liste,K.R. Steffensen, S. Theofilopoulos, C.L.Parish, C. Hazenberg, L. Ährlund Richter,O. Hovatta, J. Gustafsson, and E. Arenas.Liver X Receptors and oxysterols promoteventral midbrain neurogenesis in vivo andin human embryonic stem cells. Cell StemCell. Oct 2;5(4):409-19. (2009).R. B. Tinsley, C. Bye, C.L. Parish, A. Tziotis-Vais, S. George, J.G. Culvenor, Q. Li, C.L.Masters, , D.I. Finkelstein, M. K. Horne.Dopamine D2 receptor knockout micedevelop nigral synucleinopathy. Annals ofNeurology. 66(4):472-84 (2009).E. Arenas, C.L. Parish, I. Liste, D. Ribeiro,E.R. Andersson, C. Ramirez, V. Bryja.Enhanced dopaminergic differentiationof neural and embryonic stem cells byWnt5a, Human Gene Therapy, Vol20 (11),pp 1368-70. (2009)Harman AN, Kraus, M, Bye CR, BythK, Turville SG, Tang O, Mercier SK, Nasr,N, Stern JL, Slobedman B, DriessenC, Cunningham AL. HIV-1 InfectedDendritic Cells Show Two Phases of GeneExpression Changes With LysosomalEnzyme Activity Decreased Duringthe Second Phase. Blood. 2009 Jul2;114(1):85-94.Maddocks S, Scandurra GM, NourseC, Bye CR, Williams RB, SlobedmanB, Cunningham AL, Britton WJ. Geneexpression in HIV-1/Mycobacteriumtuberculosis co-infected macrophagesis dominated by M tuberculosis.Tuberculosis 2009 Jul;89(4):285-93.Figure 1Figure 3Figure 4Figure 2Figure 1: Wnts display highly regulated temporaland special expression in the developing brain,suggestive of a role in DA axon guidance. Sagittalview (embryonic day 13.5 mouse) of Wnt7aexpression. Note Wnt7a expression (red) adjacentto the DA fibre bundle (green), suggestive of a rolein guiding DA axonal growth towards its targets.Figure 2: Wnt5a promotes the growth andguidance of DA axons in culture.Figure 3: Dopamine neurons in an intra-striatalgraft of a PD mouse. The various stains (colours)can identify different subpopulations of dopamineneurons within the graft.Figure 4: Horizontal view of mouse brain.Grafted human embryonic stem cells are capableof restoring long distance pathways in the hostbrain.* indicates the graft site. (B) Illustratesfibres coursing through the brain alongappropriate pathways.77

Neurodegeneration— NeurodegenerationResearch OverviewThe broad interest of our group relate toParkinson’s Disease (PD), Motor NeuronDisease (MND) and the role of dopamineon frontal lobe function. There are fourgeneral themes running the research:The pathophysiology of PD and MND,Cell therapies (CT) for PD, the role ofdopamine in fore brain function and theuse of scaffolds for brain repair.The pathophysiology of PD and MND.PD and MND are both characterised byabnormal aggregation of protein in thecell. Our research in these two conditionsis travelling parallel paths. In MND thefocus is on the role of aberrant proteinsin ER stress, the movement of vesiclesand other cargo in the cell and secretionof SOD1 and related proteins. In PD, theinterest is also on secretion and howabnormal secretion acts as a marker ofdisease and also points to the locus ofpathology. Work related to this aim aredetailed below.Cell therapies for PD. Our interest hererelates to how the new dopamine cellsare recruited into brain circuitry and whatfactors regulate this. Our interest are inboth the well recognised midbrain-striatalcircuitry as well as in the pathway form theSub Ventricular Zone to the olfactory Bulb.The role of dopamine in fore brainfunction. We have shown that whendisabling movement disorders such asdyskinesias occur in PD, the dopaminelevels in the striatum are similar to thoseproduced by cocaine. Our hypothesesis that the dyskinesias are producedby the same molecular and structuralchanges as those that cause addiction.We are examining Cell therapy tools forameliorating dyskinesias and believe thatthe insights and strategies gained formstudying these movement disorders willbe relevant to addiction.The use of scaffolds for brain repair.In collaboration with the MaterialsEngineers at Monash University (JohnForsythe), we are examining how touse artificial scaffolds to improve cellsurvival, axon growth and appropriatecell connectivity. These will be used asimproved tissue culture media as wellas transplantation devices.Major Research Projectsα-synuclein in human plasmaRogan Tinsley, Xiu Xian Chia,Katya Kotchet and Malcolm Hornein collaboration with Phillip Nagley,Monash University, Gerald Shaw,University of Florida and Myles Prince,Peter MacCallum Hospital.α-synuclein is a key protein in thepathogenesis of Parkinson’s Disease (PD).It is the predominant protein in LewyBodies, which are intraneuronal inclusionsfound in the substantia nigra in PD.Mutations in the α-synuclein gene alsocause heritable forms of PD. It is likelythat obtaining a better understandingof the functions of α-synuclein in healthyand pathological neurons would lead toinsights into the pathological process ofPD as well as highlighting therapeuticpossibilities. Recently it was discoveredthat α-synuclein is secreted, and canbe found in both cerebrospinal fluidand blood plasma. We found that thelevels of circulating α-synuclein in PDand age-matched controls is different.Using Western Blotting we found thatthe α-synuclein monomer is lower inParkinson’s Disease subjects than in agematched controls. We then developed anew antibody for α-synuclein and foundthat total α-synuclein was high in peoplewith Parkinson’s Disease, even though themonomer was low, suggesting abnormalsecretion in Parkinson’s Disease. About40% of people with Parkinson’s Diseasehave α-synuclein levels that are higherthan the 90th percentile of the normalpopulation. These patients have a worseform of Parkinson’s Disease than thosewith low levels.The amount of α-synuclein found inplasma is too high to be from brainvia CSF and as blood cells expressα-synuclein, they are a potential source.We have examined α-synuclein levelsin people undergoing bone marrowablation of blood cell cancers. In thesepeople α-synuclein level in plasmafell by 50% when their peripheralblood mononuclear cells were lowest,suggesting that they make at least50% of plasma α-synuclein. When weexamined some of these blood cells wefound that they have a shorter survivalthan the same cells from control subjects.We argued therefore if the levels ofα-synuclein in the blood are altered, itsuggests that PD has interfered with thesecretion of α-synuclein by these cellsand that it may also be affecting theirlongevity. This suggests that the sameprocess affecting α-synuclein secretionin blood may also be affecting neuronesand their survival. Our current studiesare directed toward understanding howand why α-synuclein is secreted and howthis may affect neuronal function. Thelong term aim is to understand whatthe change in α-synuclein level can tellus about the disease process. We arealso interested in whether α-synucleinlevels can be used as a risk marker forParkinson’s Disease especially in peoplewith presymptomatic disease. Thesefindings may lead to biomarkers and tohigh through put screens for drugs to beused in Parkinson’s Disease.MND-linked misfolded proteinsin exosomesBrad Turner, Julie Atkin, Malcolm Hornein collaboration with Kevin Talbot,University of Oxford and Andrew Hill,University of MelbourneMND is characterised by paralysiswhich begins focally and disseminates,suggesting a spread of degenerationin the nervous system. We recentlyfound three proteins pivotal to MNDpathogenesis - SOD1, TDP-43 and FUSin exosomes. These are small secretoryvesicles derived from endosomes andreleased by cells. These exosomes wereisolated from motor neuronal culturesand rodent cerebrospinal fluid. Sincethese proteins form core components ofpathologic inclusions found in MND, theirsecretion by exosomes may contributeto disease propagation in sporadic MND.The finding of exosome-associatedTDP-43 and FUS is also surprising sincethese are DNA/RNA binding proteins,suggesting novel extracellular functionsfor these conventional nuclear proteins.There is reduced accumulation ofthese proteins in neuronally-derivedexosomes is reduced when mutationsin SOD1, TDP-43 and FUS linked toinherited MND are present. There wasan associated defect in the function ofthe early endosome in our cell models ofinherited MND. This points to abnormalprotein secretion in MND and whenconsider in the context of our previousdiscovery of endoplasmic reticulum (ER)78

stress and ER-Golgi transport failurein MND models are argues that motorneurons may be particularly susceptibleto vesicle trafficking dysfunction as partof the pathogenesis of MND. Our currentstudies are aimed at deciphering thesequence of ER, Golgi and endosomeabnormalities in MND to betterunderstand pathological pathways anddefine new therapeutic targets. We arealso addressing whether altered exosomesecretion of SOD1, TDP-43 or FUSoccurs in MND patients and correlateswith disease severity to determine thepotential biomarker value of exosomes.ER to Golgi trafficking is disrupted inMotor Neuron DiseaseJulie Atkin, Manal Farg, Adam Walker,Brad Turner, Malcolm Horne incollaboration with Paul Gleeson Universityof Melbourne and Prof Philip Nagley,Monash UniversityWe were one of the first laboratories todemonstrate that endoplasmic reticulum(ER) stress occurs early and is a keypoint in the pathophysiological cascadethat causes MND. Proteins synthesisedin the ER are transported to the Golgiapparatus for transport to other parts ofthe cell. When transport of proteins fromthe ER to Golgi fails, the Golgi becomesfragmented, leading to ER stress. Withtheir long axons, cellular transport iscrucial in motor neurons which bear thebrunt of MND. We are proposing thatthe common and early event in MNDis aberrant binding of MND proteins toproteins involved in cellular transport(such as dynein). This will disrupt ER toGolgi transport, axonal transport andimpaired vesicle trafficking, leading tofailure of protein secretion, ER stress andconsequently cell death. In support of thisproposal, a long list of vesicle, endosomeand intracellular transport proteins havebeen linked to diseases of motor neurons.Our evidence demonstrates that proteinslinked to MND: mutant SOD1, FUS andTDP-43 (but not wild-type SOD1) disruptsER to Golgi trafficking by aberrantlyto proteins such as dynein, Rab1 andCOPII. This interaction between the“MND proteins” and dynein mightdisturb efficient transport either bycompeting with other essential cargo fortransport sites or by diminishing dyneinfunction. Over-expressing a componentof dynein prevents inclusion formationand improved cellular survival. COPIIis required for proteins to exit the ER.COPII not only co-immunoprecipitateswith the “MND proteins” but it is alsoup-regulated and mis-localised intoinclusions. Rab1 is required for involvedin the docking of vesicles traffickingbetween ER and Golgi and the “MNDproteins” bind to Rab1, presumablyreducing its function. Together these datasuggest that transport from the ER to theERGIC compartment is blocked in cellsexpressing mutant “MND proteins”. Insummary, ER-Golgi trafficking is impairedin MND and proteins linked to MNDbind to proteins involved in transport,suggesting disruption to ER to Golgitransport function in MND.Figure 1Figure 2Figure 1: Fig MND proteins modulate ER toGolgi transport (A) BDNF secretion is inhibitedin cells expressing mutant SOD1, (B, C) MutantSOD1 interacts with COPII and (D) Rab 1, (E)Dynein interacts with mutant SOD1, (F) mutantTDP-43, (G) mutant FUSW=Wildtype S=Mutant SOD1 Un,GFP ControlsFigure 2: Mutant SOD1 triggers ER stress fromoutside the ER. Immunocytochemistry reveals thatmutant SOD1 A4V does not co-localise with ERmarkers ER tracker and calreticulin.79

Neurodegeneration— NeurodegenerationOther Research ProjectsStaff ListConferences and PresentationsMechanisms of DyskinesiaDavor Stanic, Doris Tomas andMalcolm HorneArtificial three dimensional scaffolds forGrowth of neurons and brain repairRogan Tinsley, Clare Parish and MalcolmHorne in collaboration with DeniseFon, Hanqing Feng and John Forsythe(Monash University) and David Finkelstein(Mental Health Research Institute)Constructing cell models ofneurodegenerative diseaseJulie Atkin, Rogan Tinsley, Brad Turner,Mohammad Gadi, Adam Walker, ManalFarg and Malcolm Horne, in collaborationwith He Ling Ng and Phillip Nagley(Monash University)Genes in Parkinson’s diseaseJustin Rubio and Malcolm Horne incollaboration with Katya Kotschett(St Vincent’s) and (Menzies ResearchCentre, Hobart)Micromixing and PCR efficiencyTim Aumann, Wah Chin Boon andMalcolm Horne in collaborationwith Richard Manasseh (CSIRO)Restoring the nigrostriatal pathwayin Parkinson’s DiseaseClare Parish, Lachlan Thompson,Doris Tomas, Malcolm HorneStem cell therapies for Parkinson’sDiseaseLachlan Thompson, Clare Parish, DorisTomas, Malcolm Horne, (collaborators:Mark Denham and Mirella Dottori, CNSUniversity of Melbourne)Fellows, Senior Research Officers, andResearch OfficersMalcolm Horne BMedSci MBBS (Hons) PhD(Monash) FRACPJulie Atkin BSc (Hons) (Manchester) Grad Dip(RMIT) PhD (Sheffield)Tim Aumann BSc (Hons) PhD (Mon)Rogan Tinsley BBiotech (Hons) PhD (Flinders)Davor Stanic BSc (Hons) PhD (Mon)Bradley Turner BSc (Hons) PhD (Melb)Katya Kotschet MBBS (Hons, Monash) FRACPResearch AssistantsManal Farg MSc (Canterbury, NZ)Doris Tomas BSc (Deakin)Hui Kheng Chua BSc (Hons)(Queensland)Kylie White BAppSc (Hons) PhD (RMIT)(from 15 Sept)Annie Hung BSc (Hons) & MSc(Queen’s University, Ontario, Canada)Ella Wilkins BSc (University of Western Australia)Hons (Melbourne)StudentsChathurini Fernando BSc MSc (Hons)(Auckland)Brette Blakely BA (Hons) (Wellesly, USA), MA(Monash)Chris Bye BSc (Hons) (UNSW)Kai Ying Soo BSc Hons (Mon) (from May 2005)Adam Walker BSc (Hons) (Tas) (from March 2006)Yi Sui BMed (Dalian, China)Jenny Chow BSc (Hons) (Monash)Jason Spencer BSc (Mon) (Masters Studentfrom July 2008)Xiu Xian Chia (AMS student) (Until May 2009)Jamie Lim (Utrecht University)Ella Wilkins7th GeneMappers Conference,speaker 16/04Adam WalkerProtein Misfolding and NeurologicalDisorders Meeting, oral presentation11/09International Symposium on ALS/MND,oral presentation 12/09Rogan TinsleyAustralian Neuroscience Society,29th Annual Meeting, Canberra. Jan 09Malcolm HorneSpeaker, A&PD Symposium SydneyMay 20093AW Dr Feelgood 27th March 2009Seminar Speaker, Eccles Institute ANUCanberra June 2009Oceania Medical School, Samoa. VisitingProfessor April 2009Griffiths University, Visiting ProfessorOct 2009Speaker, Australian Society forMovement Disorders Perth Nov 2009Julie AtkinMND Symposium, Queensland BrainInstitute. February (2010)20th International ALS/MND Symposium,Berlin, Germany December (2009)La Trobe University School of MolecularSciences June 2009Chris ByeAustralian Neuroscience Society Meeting,2009.Regulating neurogenesis in theadult brainYi Sui, Davor Stanic and Malcolm HorneAnti-inflammation therapy andParkinson’s diseaseYi Sui, Davor Stanic, Bevyn Jarrotand Malcolm Horne80

Major Collaborative LinksNationalJim StankovitchMenzies InstituteGenetics of Parkinsons diseaseProf Paul GleesonDepartment of Biochemistry,University of MelbourneSOD1 secretionProf Philip NagleyDepartment of Biochemistry,Monash Universityα-synuclein secretion, Mitochondrialfunction and apoptosis signalling inrelation to motor neuron diseaseProf Myles PrincePeter McCallum Cancer Institute,Melbourneα-synuclein secretionDr Andrew HillBIO21 Institute, Melbourne UniversityExosome function in Motor NeuroneDiseaseDr John ForsytheDepartment of Chemical Engineering,Monash UniversityBiomaterials and scaffoldsCollin PoutonMonash UniversityCell based therapies forParkinson’s DiseaseDr Phillip DicksonUniversity of NewcastleTyrosine hydroxylase phosphorylationInternationalProf Gerald ShawUniversity of Floridaα-synuclein secretionDr Bruno CauliLaboratoire de Neurobiologie etDiversité Cellulaire, ESPCI, FranceSingle cell multiplex PCRDr Yifat PrutDepartment of Physiology,Hebrew University, IsraelFunctional properties of spinocerebellarneurons in the cervical enlargement ofprimates performing voluntary movementAnders BjorklundDept. of Experimental Medical Science,Lund UniversityCell and Gene Therapy forParkinson’s diseaseDeniz KirikDept. of Experimental Medical Science,Lund UniversityCell and Gene Therapy forParkinson’s diseaseProf Tomas HökfeltDepartment of Neuroscience,Karolinska Institutet, SwedenRegulating neurogenesis in theadult brainDr Kevin TalbotUniversity of Oxford, UKProf Dame Kay DaviesUniversity of Oxford, UKProf Evan SnyderBurnham Institute for MedicalResearch, USACell based therapies forParkinson’s DiseaseAwardsAdam WalkerNina Buscombe Award, Motor NeuronDisease Association of Victoria, 2009Julie AtkinNina Buscombe Award, MND Associationof Victoria 2009Figure 1Figure 1: Double-immunofluorescencephotomicrograph showing the distribution of Y1receptor (magenta)- and Y2 receptor (green)-likeimmunoreactivity at the level of the hippocampalformation. Here, Y1 and Y2 receptor expressionwas found to surround the neurogenic subgranularzone of the dentate gyrus, where neurogenesiscontinues throughout adulthood .Figure 2Figure 2: Double immunofluorescencephotomicrograph showing doublecortin (DCx)-likeimmunoreactivity (LI) (magenta) and Y1 receptor-LI (green) in the dentate gyrus of an adult mousebrain. This is a region of the brain in which thebirth of new cells continues in the adult brain.Here we show a close association, and is somecases overlap (white) between immature neuronslabelled with doublecortin and the Y1 receptor,which is a regulator of neurogenesis in theadult brain.81

Neurodegeneration— NeurodegenerationFigure 3Figure 3: Triple-immunofluorescencephotomicrograph displaying the distribution ofneuropeptide Y- (blue), Y1 receptor- (magenta)and Y2 receptor- (green) like immunoreactivityat the level of the mouse paraventricularhypothalamic nucleus and suprachiasmaticnucleus. Dividing cells and immature neurons arepresent in this region of the brain, and we areexamining whether neuropeptide Y plays a role inregulating neurogenesis in this region of the brain.PublicationsWilkinson DJ, Kahane G, Horne M,Savulescu J. Functional neuroimagingand withdrawal of life-sustainingtreatment from vegetative patients.J Med Ethics.35:508-511 (2009)Tinsley RB, Bye CR, Parish CL et al.Dopamine D2 receptor knockout micedevelop features of Parkinson disease.Ann Neurol.66:472-484 (2009)Sui Y, Stanic D, Tomas D et al. Meloxicamreduces lipopolysaccharide-induceddegeneration of dopaminergic neuronsin the rat substantia nigra pars compacta.Neurosci Lett.460:121-125 (2009)Sritharan A, Egan G, Johnston L et al.A longitudinal diffusion tensor imagingstudy in symptomatic Huntington’sdisease. J Neurol NeurosurgPsychiatry(2009)Soo KY, Atkin JD, Horne MK, NagleyP. Recruitment of mitochondria intoapoptotic signaling correlates withthe presence of inclusions formedby amyotrophic lateral sclerosisassociatedSOD1 mutations. JNeurochem.108:578-590 (2009)Nisbet DR, Rodda AE, Horne MK et al.Neurite infiltration and cellular responseto electrospun polycaprolactonescaffolds implanted into the brain.Biomaterials.30:4573-4580 (2009)Nisbet DR, Rodda AE, Finkelstein DIet al. Surface and bulk characterisationof electrospun membranes: problemsand improvements. Colloids Surf BBiointerfaces.71:1-12 (2009)Nisbet DR, Forsythe JS, Shen W et al.Review paper: a review of the cellularresponse on electrospun nanofibers fortissue engineering. J Biomater Appl.24:7-29 (2009)Loesch DZ, Khaniani MS, Slater HR etal. Small CGG repeat expansion allelesof FMR1 gene are associated withparkinsonism. Clin Genet.76:471-476(2009)Horne MK, Nisbet DR, Forsythe JS,Parish C. Three dimensional nanofibrousscaffolds incorporating immobilizedBDNF promote proliferation anddifferentiation of cortical neural stem cells.Stem Cells Dev(2009)Horne M. Monash Bioethics Review.Vol. 28, 2009:12.11-12.12Crompton KE, Forsythe JS, Horne MK etal. Molecular level and microstructuralcharacterisation of thermallysensitive chitosan hydrogels. SoftMatter.5:4704–4711 (2009)Corben LA, Delatycki MB, Bradshaw JL etal. Impairment in motor reprogrammingin Friedreich ataxia reflecting possiblecerebellar dysfunction. J Neurol(2009)Wilkins E as part of the Australia andNew Zealand Multiple Sclerosis GeneticsConsortium (ANZgene). Genomewideassociation study identifies newmultiple sclerosis susceptibility loci onchromosomes 12 and 20. Nat Genet.2009 Jul; 41(7):824-8. Epub 2009 Jun 14J Faijerson, A Thorsell, J Strandberg,E Hanse, M Sandberg, PS Erikssonand RB Tinsley* (2009). Adult neuralstem/progenitor cells reduce NMDAinducedexcitotoxicity, via the novelneuroprotective peptide pentinin.Journal of Neurochemistry. 109(3): 858-866.Harman AN, Kraus, M, Bye CR, BythK, Turville SG, Tang O, Mercier SK, Nasr,N, Stern JL, Slobedman B, DriessenC, Cunningham AL. HIV-1 InfectedDendritic Cells Show Two Phases of GeneExpression Changes With LysosomalEnzyme Activity Decreased Duringthe Second Phase. Blood. 2009 Jul2;114(1):85-94Maddocks S, Scandurra GM, NourseC, Bye CR, Williams RB, SlobedmanB, Cunningham AL, Britton WJ. Geneexpression in HIV-1/Mycobacteriumtuberculosis co-infected macrophagesis dominated by M tuberculosis.Tuberculosis 2009 Jul; 89(4):285Walker AK, Turner B J, Atkin J DEndoplasmic Reticulum Stress andProtein Misfolding In AmyotrophicLateral Sclerosis Protein MisfoldingDisorders (2009)82

Neurodegeneration— Steroid NeurobiologyMajor Research ProjectsOther Research ProjectsStaffThe Neuroprotective EffectsOf OestrogensHui Kheng Chua, Wah Chin Boon,in collaboration with Rachel Hill,Margaret Jones and Evan Simpsonin Prince Henry’s InstituteOestrogens are converted fromandrogens in local regions of the brainby the enzyme Aromatase and signallocally through the oestogen receptorsERα and ERß. It has been difficult to studythe direct effects of sex hormones onbrain function until the development ofthe Aromatase Knockout (ArKO) mousewhich is completely estrogen-deficient.We previously detected apoptosis in thedopaminergic cells in the hypothalamusof 1 year-old male ArKO mice. Theneuroprotective effects is even morepronounce in female animals – 12 weekoldfemale ArKO mice already startedto display spontaneous apoptosis ofpyramidal neurons in the frontal cortexwhile wild-type (WT) littermates showno signs of apoptosis. Concomitantly,bcl-2 related anti-apoptotic genesare down-regulated whereas the proapoptoticgene TRADD is up-regulatedin the female ArKO frontal cortex.This phenotype can be rescued by3-week replacement of 17ß-estradiol.Furthermore, the apoptosis phenotypeis exacerbated in 12–15 month-old femaleArKO mice, which have 30% less neuronsin the frontal cortex and lower brainweights than WT counterparts. Thesedata show that estrogens are essentialfor the survival of female cortical neuronseven in the absence of pathologicalconditions or external assaults. Ourobservations also demonstrate thesexually dimorphic susceptibility ofneurons to estrogen deficiency.Oestrogens and metabolismJenny Chow, Wah Chin Boon,in collaboration with Evan Simpson (PHI)Oestrogens in diabetes developmentMichelle van Sinderen, Wah Chin Boon,in collaboration with Evan Simpson (PHI)Micromixing and PCR efficiencyTim Aumann, Wah Chin Boon andMalcolm Horne in collaboration withRichard Manasseh (CSIRO)Oestrogens and microRNAWah Chin Boon, Michelle van Sinderen,in collaboration with Colin Clyne (PHI)Oestrogens in FibrosisChrishan Samuel and Wah Chin Boon,in collaboration with Tim Hewitson(Royal Melbourne Hospital)Figure 1: Immunohistochemistry of cortex of12 week-old WT female mice Androgen Receptor(AR) and ERß are localised in the nucleus whereasERα is membrane bound. DAPI stains nuclei blue.Figure 1Senior Research OfficerWah Chin BoonResearch AssistantHui Kheng Chua BSc (Hons) (Queensland)StudentsJason Spencer BSc (Mon)Jenny Chow BSc (Hons) (Monash)Michelle van Sinderen(Hons) (Deakins)83

Neurodegeneration— Steroid NeurobiologyConferences and PresentationsCollaborationsAwardsWah Chin BoonSymposium speaker, World GeneCongress, Foshan, China, December1-7,2009Behavioural Neuroscience Meeting,Melbourne, Speaker, July 10Seminar Speaker, Mindful, Centre forTraining and Research in DevelopmentalHealth, Department of Psychiatry,University of Melbourne, 25 August 2009Seminar Speaker, School of Psychology,Psychiatry and Psychological Medicine,Monash University, 15 Sept 2009Seminar Speaker, Department ofPharmacology, Latrobe University(Bendigo Campus), 13 May 2009Michelle van SinderenOral presentation, 45th Annual Meetingof the European Association for theStudy of Diabetes, Vienna Austria29 Sept-2 Oct 2009Allan E. HerbisonDepartment of Physiology, School ofMedical Science University of Otago,New ZealandProject: Oestrogen and kisspeptinsMaarten van den BuuseMental Health Research Instituteof VictoriaTitle: Oestrogen and behaviorQiao-Xin LiDepartment of Pathology,The University of MelbourneTitle: NMDA receptors, apoptosisand neurodegenerationEvan SimpsonPrince Henry’s InstituteProject: Roles of oestrogens in obesityand diabetesJeffery ZajacDepartment of Medicine, Austin Health,University of MelbourneProject: Roles of androgens in the brainand on behaviourBruce Tonge and Nicole RinehartCentre for Developmental Psychiatry& Psychology, Monash UniversityProject: Roles of sex hormones in AutismMichelle van SinderenEndocrine Society Australia//IPSENInternational Travel Grant Award 2009European Association for the Study ofDiabetes, Travel Grant Award 2009PublicationsChow JD, Simpson ER, Boon WC.Alternative 5’-untranslated first exons ofthe mouse Cyp19A1 (aromatase) gene.J Steroid Biochem Mol Biol. 2009Jul;115(3-5):115-25.Hill RA, Boon WC. Estrogens, brain, andbehavior: lessons from knockout mousemodels. Semin Reprod Med. 2009May;27(3):218-28.Clarkson J, Boon WC, Simpson ER,Herbison AE. Postnatal development ofan estradiol-kisspeptin positive feedbackmechanism implicated in puberty onset.Endocrinology. 2009 Jul;150(7):3214-20.Hill RA, Chua HK, Jones ME, Simpson ER,Boon WC. Estrogen deficiency results inapoptosis in the frontal cortex of adultfemale aromatase knockout mice. MolCell Neurosci. 2009 May;41(1):1-7.Van Sinderen ML, Boon WC, EderveenAG, Kloosterboer HJ, Simpson ER,Jones ME. The estrogenic componentof tibolone reduces adiposity in femalearomatase knockout mice. Menopause.2009 May-Jun;16(3):582-8.84

Figure 1Figure 1: (Neuropathology of motoneuronedisease) Toxic effects of inclusion formation bymutant Cu/Zn-superoxide dismutase (mSOD1) inastrocytes. Cultures when transfected with GFPtagged-mSOD1 develop inclusions in a timedependentmanner. At 72h post-transfectiontwo populations of inclusions were noted thatwere classified by apparent nature and area ofinclusions. Micro-inclusions: diffuse (A) and diffusewith dotted inclusions (B). Macro-inclusions:some clumpy inclusions (C) and extensiveinclusions (D). Scale bar = 10µm. The presenceof mSOD1 causes an extensive change in thephenotype of astrocytes as shown by GFAPimmunocytochemistry from resting, cobble-stoned(E) to predominantly stellate (F) consistent withinflammation. Scale bar = 50µm. mSOD1 alsocauses nuclear changes indicative of fragmentationand cell death: Hoechst staining reveals healthynuclei in control (G) but nuclear profiles in variousstages of fragmentation (H). Scale bar = 50 µm.Figure 2Figure 2: (Glutamate transporters) Rotterlin,an inhibitor of protein kinase C, causes extensivestructural reorganization of cultured astrocytesat 1 (B) and 24h (C) relative to untreated cells (A)as shown by cytochemistry with phalloidin forF-actin (red). Merged images show this structuralrearrangement involves redistribution of theglutamate transporter EAAT1 (green) from aneven cell surface localization (bottom left) tomore punctuate patterns (bottom middle &right). Cell nuclei (blue).Figure 3Figure 3: (Mitochondria and neuronal injury)Cortical neurones transfected with the autophagymarker GFP tagged-microtubule associatedprotein light chain 3 (GFP-LC3) display mobilizationfrom cytosol to autophagic vesicles (B) 24h aftertreatment with the oxidative stressor, hydrogenperoxide, relative to untreated cells (A). Priortreatment with 3-methyladenine (C) substantiallyinhibits this mobilization. Scale bars = 10µm.85

NeuropeptidesResearch OverviewPeptides of the relaxin family andthe protein enzyme insulin-regulatedaminopeptidase (IRAP) are the majorresearch themes of the Neuropeptidesteam.Our long standing research focus onthe relaxin peptide family and theirreceptors continued in 2009 withimportant advances in understandingpeptide structure and function led byProfessor John Wade, the role of relaxinin fibrosis by Dr Chrishan Samuel andreceptor function and signalling fromAssociate Professor Ross Bathgate’steam. Studies on the novel neuropeptiderelaxin-3 and its GPCR receptor RXFP3have uncovered their potential rolein the modulation of arousal, sleep/wake patterns, mood and memory.Relaxin-3 research is led by AssociateProfessor Andrew Gundlach, with strongcollaborative support from Johnson &Johnson Pharmaceutical Research andDevelopment (San Diego, CA). Studieson the physiological role of the enzymeknown as IRAP have opened the wayfor the development of strategies forthe treatment of memory disorders. DrSiew-Yeen Chai and her colleagues havediscovered a series of small moleculeIRAP inhibitors with memory enhancingproperties that have exciting potential foralleviating memory deficits.The Relaxin Project continued itssuccessful clinical development in 2009with ongoing Phase III trials in acute heartfailure. This is the culmination of manyyears of research by Florey scientistsinvestigating the biological role of therelaxin peptide family and is underpinnedby a strong portfolio of internationalpatents. The Phase II trial in acute heartfailure showed strong, unequivocalimprovement in heart function. Relaxinwas safe and well tolerated and therewas evidence of a protective effect onkidney function. The Phase III clinicaltrial involves 2,000 patients worldwideand will be completed in 2011. The USFood and Drug Administration (FDA)has granted “Fast Track¨ designation torelaxin as part of its program to expeditethe review of new drugs intended to treatserious or life-threatening conditions thatcan potentially address unmet medicalneeds. In an exciting development in late2009 our commercial partner, CortheraInc (San Mateo, CA, formerly known asBAS Medical) was acquired by the largepharmaceutical company Novartis basedin Switzerland.The Neuropeptides team maintainedits competitive position in the fieldinternationally during 2009 with keypapers published in the Proceedings ofthe National Academy of Sciences USA,FASEB Journal, Molecular Pharmacology,Endocrinology, Journal of ComparativeNeurology and Learning and Memory.Professor John Wade and Dr SherieMa were awarded Australian Academyof Science awards for research visits toEurope and many of Neuropeptide teamstudents received travel awards to attendNational and International meetings.The team continues to attract the bestgraduate students which this yearincluded 5 Honours students and 14 PhDscholars. Additionally the NeuropeptidesTeam had visiting scientists from Canada,France, Sweden and Korea working withmembers of the team during the year.Major Research ProjectsRole of relaxin-3 neural networksin brain revealed by studies intransgenic miceAndrew Gundlach, Craig Smith, SherieMa, Qian Sang, M Akhter Hossain, JohnWade, Ross Bathgate, in collaborationwith Steven Sutton, Christine Dugovic,James Shoblock, Timothy W Lovenberg,Neuroscience Drug Discovery, Johnsonand Johnson Pharmaceutical Researchand Development, San Diego, USAA major challenge in neuroscience is toidentify the neural circuits that underliespecific behaviours and determinehow they are perturbed in psychiatricdisorders – a task aided in recent yearsby neuroimaging studies of control andpatient groups; better animal modelsof human behaviour and psychiatricillness; and more integrated scientificinvestigations.One powerful experimental approach isto study brain chemistry and behaviourin ‘knockout’ mice in which genes forneurotransmitter-related proteins aredeleted. This has led to insights into therole of various monoamine and peptidetransmitters and their neural receptorsin arousal and circadian activity; in stressand fear responses and in mood control.And it is these complex inter-relatedbehaviours that are perturbed in clinicalconditions, such as insomnia, anxietyand depression.Anatomical and physiological studies ofnewly-identified relaxin-3/RXFP3 neuralnetworks in brain have identified theirpossible modulation of arousal andsleep/wake patterns, and memory. Thedistribution of relaxin-3/RXFP3 in rat andmouse brain also suggests interactionswith other transmitters and peptidesknown to be involved in behaviouralstate and affective behaviour. Moredetails of these interactions are required,however, and further testing is needed toestablish relaxin-3’s precise function. Incurrent studies, therefore, we are usingtransgenic mice to explore the integrativerole of relaxin-3/RXFP3 signalling.Behaviour of relaxin-3 knockoutmice and role of relaxin-3 in sleeparchitecture and moodA new map of relaxin-3/RXFP3 networksin adult mouse brain (Figure A) providesa basis for behavioural studies of twogene-deletion mouse strains – relaxin-3and RXFP3 knockout mice. Excitingpreliminary data indicates relaxin-3deficient mice have altered motor activityand sleep patterns during their normalactive (night) phase. Relaxin-3 KO miceexhibit a 50% reduction in dark-phaserunning wheel activity (Figure B) andincreased sleeping, with more frequentrapid eye movement (REM) sleepepisodes resulting in an increased REMsleep duration over the entire light-darkcycle, relative to wildtype (WT) mice.In a ‘tail suspension test’, routinelyused to assess levels of depressive-likebehaviour and antidepressant drugactivity, relaxin-3 KO and WT micedisplayed similar immobility after vehicle,but decreases in immobility time inducedby antidepressant treatment were evidentin WT, but not relaxin-3 KO mice. Thesechanges reflect hallmark symptoms ofdepression in patients and in animalmodels of the condition, and suggestthese mice may represent a new geneticmodel of clinical depression.In a test of hedonic or rewardingresponses in mice, intake of palatable,high-fat food was measured in relaxin-3KO and WT mice after naloxone orvehicle treatment. Relaxin-3 KO mice ateless palatable food than their littermates86

under basal conditions, and naloxoneinhibited palatable food intake in WT, butnot in KO mice. In contrast, WT and KOmice ate similar levels of food after fooddeprivation. These findings suggest alow hedonic tone and further implicaterelaxin-3 signalling in mood control.Studies to identify RXFP3 sensitiveneurons and circuits are currentlyunderway using RXFP3 KO mice (FigureC) and another transgenic mouse strainin which RXFP3 expression is reflectedin nerves by the presence of a greenfluorescent protein (eGFP; Figure D).Figure 1: (A) Map of relaxin-3 and RXFP3 atthe level of ventral hippocampus in the mouse.(B) Relaxin-3 KO mice exhibit a near 50%reduction in running wheel activity and increasedsleeping; with more frequent REM sleep duringthe dark phase, relative to WT mice. In (C)RXFP3 KO and (D) RXFP3 BAC-eGFP mousestrains, neuronal RXFP3 expression is indicatedby the presence of an enzyme or a bright greenfluorescent protein, respectively.Figure 1AFigure 1BFigure 1CFigure 1DRXFP3 KO/LacZ KITSBAC RXFP3-GFP87

NeuropeptidesDesign and development of relaxin-3analogues capable of crossing theblood brain barrier (BBB)M Akhter Hossain, Fazel Shabanpoor,Andrew Gundlach, Ross Bathgate andJohn Wade in collaboration with Dr JohanK. Rosengren, Institute of MolecularBiology, University of QueenslandHuman relaxin-3 (H3 relaxin), the mostrecently discovered member of therelaxin superfamily that was discoveredby our laboratory in 2002, has emerged asa key brain neuropeptide; and H3 relaxinanalogues have considerable clinicalpotential for the treatment of variousdisorders such as obesity, anxiety, mood,sleep disorders and memory dysfunction.Like insulin, H3 relaxin is composed of acomplex two chain structure that is linkedby three disulfide bonds. Surprisinglythough, it binds and activates all fourof the relaxin family peptide receptors(RXFP1-4), despite having the highestaffinity for its native receptor, RXFP3. Thus,the potential therapeutic use of H3 relaxinis complicated by this receptor crossreactivity.Recently, however, we haveidentified the key residues in the B-chainof H3 relaxin that define its specificity forRXFP3. This information, together withour determination of the NMR solutionstructure of H3 relaxin, has allowed us todesign RXFP3 receptor specific agonistsand antagonists. Over the past 12-months,we have generated and characterizedtwo lead compounds, an agonist and anantagonist, that are RXFP3 specific andmuch smaller and simpler compared tonative H3 relaxin. Our current focus is toutilize these newly generated analoguesto design and chemically synthesizeadditional domain minimized andproteolytically-stable analogues of H3relaxin. These small but stable analoguesare better expected to cross the BBB toelicit specific biological actions. To assistthem to cross the BBB, strategies aimedat conjugating them to small carriermolecules (such as Lys-palmitoyl or SynB3vector peptide) will also be carried out.These peptidomimetics, with improvedpharmacokinetic properties, will be keylead molecules for future pharmaceuticaldevelopment.We are using state-of-the-art peptidesynthesis technology and drug designtools in conjunction with NMR techniques,to prepare proposed H3 relaxinanalogues in sufficient quantities to allowtheir subsequent detailed biological study,using molecular biology, pharmacologyand neurobiology techniques.Other Research ProjectsInvestigating the role of relaxin asan anti-fibrotic therapyChrishan Samuel, Chongxin Zhao, SuEe Wong, Bryna Chow, Ross Bathgateand Geoffrey Tregear in collaborationwith Associate Professor Xiao-Jun Du,Baker Heart Research Institute, AssociateProfessor Tim Hewitson, Royal MelbourneHospital, Professor Roger Summers,Monash University and Associate ProfessorMimi Tang, Murdoch Children’s ResearchInstitute, Royal Children’s HospitalWe have previously shown that humangene-2 (H2) relaxin (the major storedand circulating form of human relaxin,which is bioactive in rodents) is bothan endogenous inhibitor of collagendeposition and a rapidly-acting butsafe therapy for the progressive fibrosis(scar tissue) that occurs during aberranttissue repair and end-stage organdisease. The anti-fibrotic actions ofH2 relaxin have been demonstratedin diverse experimental models ofcardiovascular, renal, pulmonary/airway,dermal and hepatic disease and areprimarily mediated via its ability toinhibit transforming growth factor beta-1(TGF-ß1; a major pro-fibrotic cytokine)-stimulated myofibroblast differentiationand collagen production.We recently employed primary ratrenal cortical and cardiac ventricularmyofibroblast cultures in vitro, in additionto mouse models of tubulointerstitialkidney/interstitial heart disease in vivo(in which TGF-ß1 was up-regulated), tofurther delineate the signal transductionpathways involved with the ability of H2relaxin to disrupt TGF-ß1 activity. H2relaxin was found to signal through itsprimary receptor, Relaxin Family PeptideReceptor 1 and the nitric oxide (NO)pathway (specifically via neuronal nitricoxide synthase (nNOS), NO and cGMP)to down-regulate the phosphorylationof Smad2 (a regulatory protein thatpromotes the actions of TGF-ß1); as ameans of down-regulating TGF-ß1 activityand hence, the influence of TGF-ß1 onmyofibroblast differentiation and collagendeposition. Our ongoing research aims tofurther explore other signal transductionpathways that are associated withthe relaxin-NO interaction; as furtherknowledge of the mechanisms of H2relaxin’s anti-fibrotic actions, particularlyat the cell signalling level, will enhanceour ability to identify novel targets thatwill improve its therapeutic potential.Additional Research ProjectsFunctional and structural analysis ofRelaxin Family Peptide ReceptorsRoss Bathgate, Roy Kong, NatalieWitteveen, Daniel Scott, Tania Ferraro,Sharon Layfield in collaboration withEmma Petrie, Patrick Shilling and PaulGooley, Department of Biochemistry andMolecular Biology, University of MelbourneViral strategies for studying relaxinfamily peptide functionGabrielle Callander, Despina Ganella,Andrew Gundlach and Ross Bathgatein collaboration with Professor WalterThomas, University of QueenslandRelaxin Family Peptide ReceptorsignalingRoss Bathgate and Sharon Layfieldin collaboration with Michelle Halls,Martina Kocan and Roger Summers,Monash UniversityRole of IRAP in cardiovascular controlSiew Yeen Chai, Anthony Albiston andPeta Burns in collaboration with AssociateProfessor Robert Widdop and Dr TraceyGaspari, Department of Pharmacology,Monash UniversityCharacterisation of the global andtissue specific IRAP Knockout mouseSiew Yeen Chai, Anthony Albiston, PetaBurns, Shanti Diwakarla, Leelee Ng, ViPham and Holly Yeatman in collaborationwith Professor Yvette Michotte, ProfessorIlse Smolders and Dr Patrick Vanderheyden,Vrije University, Brussels, BelgiumRole of IRAP in ischemic brain damageSiew Yeen Chai, Anthony Albiston, PetaBurns, Shanti Diwakarla and Vi Pham incollaboration with Dr David Howells88

Development of small moleculeinhibitors of IRAPSiew Yeen Chai, Anthony Albistonand Leelee Ng in collaboration withProf Michael Parker, St Vincent’s Instituteof Medical Research and Dr PhillipThompson, Faculty of Pharmacyand Pharmaceutical Sciences,Monash UniversityRole of IRAP in Alzheimer’s diseaseSiew Yeen Chai, Anthony Albiston andHolly Yeatman in collaboration withDr Anthony White, Department ofPathology, University of MelbourneProtein profiling of kidney fibrosisand its treatment with relaxinEleni Giannakis, Chrishan Samuel,Geoffrey Tregear and John Wade incollaboration with Dr Tim Hewitson,Department of Nephrology, RoyalMelbourne HospitalIdentification of biomarkers inParkinson’s Disease and their use intherapeutic interventionEleni Giannakis, Geoffrey Tregearand John Wade in collaboration withDr Rogan Tinsley and Professor MalHorne, Howard Florey InstituteRole of relaxin-LGR7 signallingin mouse brain – insights from areceptor knockoutAndrew Gundlach, Ross Bathgate,Pei-Juan Shen, Loretta Piccenna, incollaboration with Professor AndrewLawrence, Neural Injury and RepairGroup and Professor Michael McKinley,Neurobiology of Homeostasis Group,Howard Florey InstituteRole of INSL3/LGR8 signalling insensorimotor functionAndrew L Gundlach, Qian Sang,Katayoun Sedaghat, Nilushi Karunaratne,in collaboration with David I Finkelstein,Mental Health Research Instituteof VictoriaRole of galanin in adult neural stemcells and in oligodendroglial function inhealth and diseaseAndrew Gundlach, Pei-Juan Shen, incollaboration with Dr Helmut Butzkuevenand Professor Trevor Kilpatrick, MultipleSclerosis Group, Howard Florey Institute,Dr Ann Turnley, Centre for Neuroscience,University of Melbourne, and Dr Ji Maand Professor Chong-Gang Yuan, EastChina Normal University, Shanghai, ChinaDrug design and development of newgeneration antibacterial agents basedon insect antimicrobial peptidesFeng Lin and John Wade in collaborationwith Professor Laszlo Otvos, SbarroInstitute, USA, and Professor RalfHoffmann, University of Leipzig, GermanyInvestigating the therapeutic potentialof relaxin in experimental diabetesChrishan Samuel and Su Ee Wong incollaboration with Assoc. Professor TimHewitson, Department of Nephrology,Royal Melbourne Hospital and Assoc.Professor Darren Kelly, Department ofMedicine, St. Vincent’s HospitalConformational analyses ofamyloid-beta peptides boundto lipid membranesFazel Shabanpoor and John Wade incollaboration with Professor FrancesSeparovic, Department of Chemistry,University of MelbourneInvestigating the electrophysiologyof relaxin-3/nucleus incertus neuronsSherie Ma and Andrew Gundlach incollaboration with Dr Anthony Verberne,Department of Medicine (AH/NH),University of MelbourneRole of relaxin-3 in emotional andspatial memorySherie Ma and Andrew Gundlach incollaboration with A/Prof FranciscoOlucha-Bordonau, Department of HumanAnatomy and Embryology, University ofValencia, Spain and A/Prof Angel Nunez,Department of Anatomy, Histology andNeuroscience, University AutonomaMadrid, SpainStaff ListFellows, Senior Research Officers, andResearch OfficersGeoffrey Tregear AM, BSc (Melb) PhD HonDSc(Mon) FAA, FRACIAnthony Albiston BSc (Hons) (Melb) PhD (Mon)Ross Bathgate BSc (Hons) PhD (Qld)David Bowser BSc (Hons) Grad Dip Info Tech(RMIT) PDM PhD (Melb)Siew Yeen Chai BSc (Hons) (Mon) PhD (Melb)Shanti Diwakarla BSc (Hons) PhD (Melb)Eleni Giannakis BSc (Hons) PhD (Flinders)Andrew Gundlach BSc (Hons) DipEd (Mon)PhD (Melb)M. Akhter Hossain BSc (Hons) MSc (Dhaka)PhD (Tokyo)Sherie Ma BSc (Hons) (Mon) PhD (Melb)Tomris Mustafa BSc (Hons) (VUT) PhD (Melb)Vi Pham BSc (Hons) PhD (Melb)Chrishan Samuel BSc (Hons) (Mon) PhD (Melb)Qian Sang MD (Beijing) PhD (Melb)Pei-Juan Shen BSc (Hons) (East China Normal)PhD (Mon)Maria Shleper BSc (Hons) PhD (Technion)(to October 2009)Craig Smith BBSc BSc (Hons) PhD (Melb)(from September 2009)Ernesto Vargas BSc (Hons) (Colombia)PhD (Melb)John Wade BSc (Hons) PhD (Mon) CSci CChemFRACI FRSCSuode Zhang BSc MSc PhD (Peking)Research AssistantsPeta Burns BSc (Melb)Lynley Cordeiro BSc (Hons) (Mon)Tania Ferraro BAppSc (SUT)Sharon Layfield BSc (Hons) (La Trobe)MSc (Melb)Feng Lin BSc (Hons) (Shanghai)Leelee Ng BBSc BSc (Hons) (Melb)Chongxin Zhao BSc (China) MSc (Melb)StudentsAvantika Banerjee BBSc BSc (Hons) (Melb)Alessia Belgi BSc MSc (Padova)89

NeuropeptidesGabrielle Callander BBSc BSc (Hons) (Melb)Linda Chan BSc (Hons) (Melb) (from July 2009)Bryna Chow BBSc (Melb) (to November 2009)Despina Ganella BBSc BSc (Hons) (Melb)(from March 2009)Nilushi Karunaratne BSc (Melb)(to November 2009)Chze Khai Roy Kong BSc (Hons) (Melb)Seyoung Lee BSc (Melb) (to November 2009)Broden Morgan BSc (Melb)Nicholas Ng BSc (Melb) (to November 2009)Loretta Piccenna BSc (Hons) (Mon)(to April 2009)Philip Ryan MBBS (Hons) BMedSc (Melb)Fazel Shabanpoor BSc (Hons) (Melb)Craig Smith BBSc BSc (Hons) (Melb)(to August 2009)Maneesha Weerasinghe BSc (Melb)(to November 2009)Natalie Witteveen BSc (Hons) (Melb)Su Ee Wong BBSc (Hons) (Melb)Holly Yeatman BSc (Hons) (Murdoch University)Visiting ScholarsAssoc Prof Derek Lobb, Sabatical,McMaster University, CanadaCédric Peirs, MSc student(University of Bordeaux II)Hanna Kastman, BPh student(Uppsala University)Seon-Yeong Kwak, PhD student(Seoul National University)Conferences andPresentationsRoss Bathgate5th International Symposium onPeptide Receptors, Quebec City,Canada (invited speaker)Australian Society for PsychosocialObstetrics and Gynecology, Sydney,NSW (invited speaker)Veterinär-Physiologisch-ChemischesInstitut, Leipzig, Germany (invitedseminar)Siew Yeen ChaiDepartment of ExperimentalNeuropharmacology, Vrije University,Brussels, Belgium (invited seminar)The renin-angiotensin system workshop,Monash University (invited speaker)Shanti DiwakarlaAustralian Neuroscience Society AnnualConference, Sydney, NSWAndrew GundlachEuropean Neuropeptide Club (ENC)and Summer Neuropeptide Conference(SNP) – Neuropeptides Festival, Salzburg,Austria (invited speaker)Neuropharmacology ‘NeuropeptidesSymposium’, Chicago, Illinois, USASociety for Neuroscience Annual Meeting,Chicago, Illinois, USAKarolinska Institute, Stockholm, Sweden(invited seminar)M. Akhter Hossain2nd Modern Solid Phase PeptideSynthesis and Its Applications Symposium,Gold Coast, Queensland (invited speaker)8th Australian Peptide Conference,Couran Cove, Queensland4th Asia-Pacific Peptide Symposium,Jeju Island, Korea (invited speaker)Nilushi KarunaratneAustralian Neuroscience Society,Sydney, NSWChze Khai Roy KongPost-graduate student Award Session;ASCEPT Annual Scientific Meeting,Sydney, NSWSherie Ma2nd Biennial Meeting of the InternationalSociety for Neurochemistry, Busan,South Korea (invited speaker)Vi Pham32nd Annual Meeting of the JapanNeuroscience Society, Nagoya, JapanPhil RyanSociety for Neuroscience Annual Meeting,Chicago, Illinois, USAChrishan SamuelKeystone Symposium on Fibrosis,Keystone, Colorado, USANational Heart Foundation Conference,Brisbane, Queensland (selected speaker)Bernard O’Brien Institute of Microsurgery,Melbourne, Victoria (invited seminar)Fazel Shabanpoor2nd Modern Solid Phase PeptideSynthesis and Its Applications Symposium,Gold Coast, Queensland (selectedspeaker)33rd Annual Conference of the AustralianSociety for Biophysics, Ballarat, Victoria(selected speaker)21st American Peptide Symposium,Bloomington, Indiana, USA8th Australian Peptide Conference,Couran Cove, QueenslandCraig SmithSociety for Neuroscience Annual Meeting,Chicago, Illinois, USAAwardsGabrielle CallanderHarold Mitchell Travel Award (for best 3rdyear student presentation at HFI)Despina GanellaBest First Year Speaker, Dept ofBiochemistry and Molecular BiologyPostgraduate RetreatM. Akhter HossainFNI Travel AwardSherie MaInternational Society for NeurochemistryYoung Investigator AwardAustralian Academy of Science/FrenchEmbassy of Australia Scientific Visits toEurope AwardBroden MorganDean of Science Honour’s list for the topHonours student in Physiology for 2009Vi PhamJapan Neuroscience Society Travel Award90

Phil RyanDowd ScholarshipUniversity of Melbourne, MelbourneAbroad Travelling ScholarshipFazel ShabanpoorMarch of Dimes Travel AwardStudent Award, 33rd Annual Conferenceof the Australian Society for BiophysicsYoung Investigator Travel Award, 8thAustralian Peptide ConferenceDavid Hay Post-graduate Write-up Award,University of MelbourneJohn WadeAustralian Academy of ScienceInternational Linkages-ScienceAcademies Travelling FellowshipMajor Collaborative LinksNationalProf David Craik and Dr JohanRosengrenInstitute of Molecular Bioscience,University of QueenslandTertiary structure determinationof relaxin and INSL3 peptidesDr Xiao-Jun DuBaker Heart Research Institute,MelbourneRole of relaxin in the heart andpotential of relaxin as a treatmentfor cardiac fibrosisDr Marie GibbsDepartment of Anatomy and Cell Biology,Monash UniversityThe role of astrocytes in learningand memory formationA/Prof Paul GooleyDepartment of Biochemistry andMolecular Biology, University ofMelbourneStructural studies on the relaxin familypeptide receptorsDr Tim HewitsonDepartment of Nephrology, RoyalMelbourne Hospital, MelbourneInvestigating the therapeutic potentialof relaxin in interstitial renal fibrosisDr Tony HughesDepartment of Pharmacology,University of MelbourneDesign and synthesis of conformationallyconstrained mimetics of relaxin and INSL3Prof John HutsonDouglas Stephens Surgical Laboratory,Murdoch Children’s Research Institute,MelbourneThe role of RLF (INSL3) in gubernaculardevelopmentProf Richard IvellSchool of Molecular and BiomedicalSciences, University of AdelaideBiology of INSL3Prof Michael ParkerSt Vincent’s Institute of Medical Research,MelbourneUnderstanding protein structure ofIRAP and its interactions with inhibitorsA/Prof Ray RogersReproductive Medicine Unit,Department of Obstetrics andGynecology, University of AdelaideThe function of ovarian follicular INSL3Prof Roger SummersDepartment of Pharmacology,Monash UniversityRelaxin receptor signalingA/Prof Mimi TangDepartment of Allergy and Immunology,Murdoch Children’s Research Institute,MelbourneInvestigating the therapeutic effectsof relaxin on airway remodelling andfunctionProf Walter ThomasSchool of Biomedical Sciences,University of QueenslandViral strategies for studying relaxinpeptide functionDr Phillip ThompsonDepartment of Medicinal Chemistry,Monash UniversityDesign of small molecule IRAP inhibitorsDr Jose VilladangosWalter and Eliza Hall InstituteRole of IRAP in antigen cross presentationDr Anthony WhiteDepartment of Pathology,University of MelbourneRole of IRAP in Alzheimer’s diseaseA/Prof Tony VerberneDepartment of Medicine – Austin andNorthern Health, University of MelbourneRelaxin-3 systems in the brain:Neurophysiology and behaviourDr Robert Widdop andDr Tracey GaspariDepartment of Pharmacology,Monash UniversityInvestigations into the cardiovascularfunction of the IRAP knockout mouseInternationalDr Edward AmentoMolecular Medicine Research Institute,San Francisco,USARelaxin and extracellular matrix biologyDr Anna Blasiak and Dr Tomasz BlasiakDepartment of Neurophysiology &Chronobiology, Jagiellonian University,Krakow, PolandRelaxin-3 systems and biorhythmsDr Marco CapagnoMRC Anatomical NeuropharmacologyUnit, Oxford, UKCellular physiology of relaxin-3 inamygdalaProf Alon ChenDepartment of Neurobiology,Weizmann Institute of Science,Rehovot, Israel (and Prof AndrewLawrence, Florey Neuroscience Institutes)Interaction of relaxin-3 and CRF systemsA/Prof Gavin DaweDepartment of Pharmacology, NationalUniversity of Singapore, SingaporeRelaxin-3 systems in adult neurogenesisand hippocampal functionProf Pierre De MeytsReceptor Systems Biology Laboratory,Hagedorn Research Institute, DenmarkDimerization of relaxin family peptidereceptors91

NeuropeptidesProf Aaron HsuehDivision of Reproductive Biology,Department of Gynecology andObstetrics, Stanford University MedicalCentre, Stanford University, CA, USADefining the structural domains ofthe LGR7 and LGR8 receptors anddetermining the functions of INSL3in the gonadsDr Patrick KehoeDepartment of Clinical Science atNorth Bristol, University of Bristol,Frenchay Hospital, Bristol, UKIRAP gene polymorphism andAlzheimer’s diseaseDr Jose LanciegoNeurosciences Division, CIMA,University of Navarra, Pamplona, SpainComparative studies of relaxin-familypeptides and receptors in non-humanprimateDr Tim LovenbergPharmaceutical Research andDevelopment, Johnson and Johnson,San Diego, USADistribution, regulation and functionof relaxin-3 and GPCR135 in the brainProf Thomas McCownDepartment of Psychiatry, University ofNorth Carolina, Chapel Hill, NC, USAViral strategies for studying relaxin-3functionDr Ewan McNaySection of Endocrinology, Yale UniversitySchool of Medicine, New Haven, CT, USARole of IRAP in modulating brain glucoseuptakeProf Yvette Michotte, Prof IlseSmolders and Dr Patrick Vanderheyden,Vrije University, Brussels, BelgiumCharacterisation of the global and tissuespecific IRAP Knockout mouseDr Sietse Mosselman andDr Anne RiesewijkDepartment of Pharmacology, NVOrganon, The NetherlandsPhenotyping the LGR7 KO mouseand cloning and characterising rodentLGR7 genesProf Angel NunezDepartment of Anatomy, Histology& Neuroscience, University AutonomaMadrid, Madrid, SpainNucleus incertus and relaxin-3in theta rhythm generationA/Prof Francisco Olucha-BordonauDepartment of Human Anatomy andEmbryology, University of Valencia,Valencia, SpainAnatomy and function of nucleus incertusand relaxin-3 pathwaysProf Lazlo Otvos JrThe Wistar Institute, Philadelphia, USAStructural analyses of relaxin and INSL3Prof David SherwoodDepartment of Molecular and IntegrativePhysiology, University of Illinois, USAStudies on rodent relaxin receptorsDr Dennis Stewart andDr Elaine UnemoriBAS Medical, San Mateo, USADevelopment of relaxin as anantifibrotic drugProf Masafumi TsujimotoLab of Cellular Biochemistry, RIKEN,Saitama, JapanCharacterisation of aminopeptidaseactivityProf Georges VanquelinDepartment of Molecular andBiochemical Pharmacology, VrijeUniversiteit Brussel, BelgiumRole of IRAP in immune cellsand on neurotransmitter releaseEditorial PositionsRoss BathgateMolecular and Cellular EndocrinologySiew Yeen ChaiOpen Enzyme Inhibition JournalAndrew GundlachJournal of Chemical NeuroanatomyFrontiers in NeuroscienceNeuroSignalsxPharm, Elsevier Science onlinePharmacology database (Contributor)Chrishan SamuelImmunology, Endocrine and MetabolicAgents in Medicinal ChemistryGeoffrey TregearPeptide ScienceProtein and Peptide LettersJohn WadeInternational Journal of Peptide Researchand Therapeutics (co-Editor-in-Chief)Journal of Peptide Science (Editor)Peptides (Editor)Protein and Peptide Letters (Editor)Chemical Biology & Drug DesignInternational Journal of Peptides92

PublicationsRefereed Journal ArticlesCallander GE, Thomas WG, BathgateRAD. (2009) Development andoptimization of microRNA againstrelaxin-3. Ann NY Acad Sci 1160:261-264.Callander GE, Thomas WG, BathgateRAD. (2009) Prolonged RXFP1 and RXFP2signaling can be explained by poorinternalization and a lack of beta-arrestinrecruitment. Am J Physiol Cell Physiol296:C1058-1066.De Bundel D, Smolders I, Yang R,Albiston AL, Michotte Y, Chai SY. (2009)Angiotensin IV and LVV-haemorphin 7enhance spatial working memory in rats:effects on hippocampal glucose levelsand blood flow. Neurobiol Learn Mem92:19-26.Du XJ, Xu Q, Lekgabe ED, Gao XM,Kiriazis H, Moore XL, Dart AM, TregearGW, Bathgate RAD, Samuel CS. (2009)Reversal of cardiac fibrosis and relateddysfunction by relaxin: experimentalfindings. Ann NY Acad Sci 1160:278-284.Feng S, Ferlin A, Truong A, Bathgate RAD,Wade JD, Corbett S, Han S, Tannour-Louet M, Lamb DJ, Foresta C, AgoulnikAI. (2009) INSL3/RXFP2 signaling intesticular descent. Ann NY Acad Sci1160:197-204.Giannakis E, Macris M, Tregear GW,Samuel CS, Wade JD. (2009) Evaluation ofrelaxin’s anti-fibrotic actions by SELDI-TOF mass spectromtery-based proteinprofiling of relaxin KO mice, a model ofprogressive fibrosis. Ann NY Acad Sci1160:350-352.Giannakis E, Samuel CS, Hewitson TD,Boon WM, Macris M, Reeve S, LawrenceJ, Smith AI, Tregear GW, Wade JD. (2009)Aberrant protein expression in plasmaand kidney tissue during experimentalobstructive nephropathy. Proteomics ClinApplic 3:1211-1224.Gundlach AL, Ma S, Sang Q, ShenPJ, Piccenna L, Sedaghat K, Smith CM,Bathgate RA, Lawrence AJ, Tregear GW,Wade JD, Finkelstein DI, Bonaventure P,Liu C, Lovenberg TW, Sutton SW. (2009)Relaxin family peptides and receptorsin mammalian brain. Ann NY Acad Sci1160:226-235.Halls ML, Hewitson TD, Moore XL, DuXJ, Bathgate RAD, Summers RJ. (2009)Relaxin activates multiple cAMP signalingpathway profiles in different target cells.Ann N Y Acad Sci 1160:108-111.Halls ML, Papaioannou M, Wade JD,Evans BA, Bathgate RAD, Summers RJ.(2009) RXFP1 couples to the Galpha-Gbetagamma-PI3K-PKCzeta pathwayvia the final 10 amino acids of thereceptor C-terminal tail. Ann NY Acad Sci1160:117-120.Halls ML, van der Westhuizen ET, WadeJD, Evans BA, Bathgate RAD, SummersRJ. (2009) Relaxin family peptide receptor(RXFP1) coupling to G(alpha)i3 involvesthe C-terminal Arg752 and localizationwithin membrane Raft Microdomains. MolPharmacol 75:415-428.Hartley BJ, Scott DJ, Callander GE,Wilkinson TN, Ganella DE, Kong CK,Layfield S, Ferraro T, Petrie EJ, BathgateRAD. (2009) Resolving the unconventionalmechanisms underlying RXFP1 andRXFP2 receptor function. Ann NY AcadSci 160:67-73.Haugaard-Jönsson LM, Hossain MA, DalyNL, Bathgate RAD, Wade JD, Craik DJ,Rosengren KJ. (2009) Structural propertiesof relaxin chimeras. Ann NY Acad Sci1160:27-30.Haugaard-Jönsson LM, Hossain MA,Daly NL, Craik DJ, Wade JD, RosengrenKJ. (2009) Structure of human insulinlikepeptide 5 and characterizationof conserved hydrogen bonds andelectrostatic interactions within the relaxinframework. Biochem J 419:619-627.Hewitson TD, Samuel CS. (2009) Relaxin:an endogenous renoprotective factor?Ann NY Acad Sci 1160:289-293.Hossain MA, Bathgate RAD, RosengrenKJ, Shabanpoor F, Zhang S, Lin F, TregearGW, Wade JD. (2009) The structural andfunctional role of the B-chain C-terminalarginine in the relaxin-3 peptideantagonist, R3(BDelta23-27)R/I5. ChemBiol Drug Des 73:46-52.Hossain MA, Bathgate RAD, Tregear GW,Wade JD. (2009) De novo design andsynthesis of cyclic and linear peptidesto mimic the binding cassette of humanrelaxin. Ann NY Acad Sci 1160:16-19.Hossain MA, Belgi A, Lin F, Zhang S,Shabanpoor F, Chan L, Belyea C, TruongHT, Blair AR, Andrikopoulos S, TregearGW, Wade JD. (2009) Use of a temporary“solubilizing” peptide tag for the Fmocsolid-phase synthesis of human insulinglargine via use of regioselective disulfidebond formation. Bioconjug Chem20:1390-1396.Hossain MA, Rosengren KJ, Zhang S,Bathgate RAD, Tregear GW, van LieropBJ, Robinson AJ, Wade JD. (2009) Solidphase synthesis and structural analysis ofnovel A-chain dicarba analogs of humanrelaxin-3 (INSL7) that exhibit full biologicalactivity. Org Biomol Chem 7:1547-1553.Liu C, Kuei C, Sutton S, Shelton J, Zhu J,Nepomuceno D, Hossain MA, Wade JD,Bathgate RAD, Bonaventure P, LovenbergT. (2009) Probing the functional domainsof relaxin-3 and the creation of a selectiveantagonist for RXFP3/GPCR135 overrelaxin receptor RXFP1/LGR7. Ann NYAcad Sci 1160:31-37.Luo X, Bathgate RAD, Liu YL, Shao XX,Wade JD, Guo ZY. (2009) Recombinantexpression of an insulin-like peptide3 (INSL3) precursor and its enzymaticconversion to mature human INSL3. FEBSJ 276:5203-5211.Ma S, Olucha-Bordonau FE, Hossain MA,Lin F, Kuei C, Liu C, Wade JD, Sutton SW,Nuñez A, Gundlach AL. (2009) Modulationof hippocampal theta oscillations andspatial memory by relaxin-3 neurons ofthe nucleus incertus. Learn Mem 16:730-742.Ma S, Sang Q, Lanciego JL, Gundlach AL.(2009) Localization of relaxin-3 in brainof Macaca fascicularis: identification ofa nucleus incertus in primate. J CompNeurol 517:856-872.Ma S, Shen PJ, Sang Q, Lanciego JL,Gundlach AL. (2009) Distribution ofrelaxin-3 mRNA and immunoreactivityand RXFP3-binding sites in the brain ofthe macaque, Macaca fascicularis. AnnNY Acad Sci 1160:256-258.Moodley Y, Atienza D, ManuelpillaiU, Samuel CS, Tchongue J, IlancheranS, Boyd R, Trounsan A. (2009) Humanumbilical cord mesenchymal stem cellreduce fibrosis of bleomycin-inducedlung injury. Am J Path 175:303-313.93

NeuropeptidesMookerjee I, Hewitson TD, Halls ML,Summers RJ, Mathai ML, Bathgate RAD,Tregear GW, Samuel CS. (2009) Relaxininhibits renal myofibroblast differentiationvia RXFP1, the nitric oxide pathway andSmad2. FASEB J 23:1219-1229.Pham V, Burns P, Albiston AL, YeatmanHR, Ng L, Diwakarla S, Chai SY. (2009)Reproduction and maternal behavior ininsulin-regulated aminopeptidase (IRAP)knockout mice. Peptides 30:1861-1865.Rosengren KJ, Bathgate RAD, Craik DJ,Daly NL, Haugaard-Jönsson LM, HossainMA, Wade JD. (2009) Structural insightsinto the function of relaxins. Ann NY AcadSci 1160:20-26.Royce SG, Miao YR, Lee M, Samuel CS,Tregear GW, Tang MLK. (2009) Relaxinreverses airway remodelling and airwaydysfunction in allergic airways disease.Endocrinology 150:2692-2699.Rozgonyi F, Szabo D, Kocsis B, OstorháziE, Abbadessa G, Cassone M, Wade JD,Otvos L Jr. (2009) The antibacterial effectof a proline-rich antibacterial peptide A3-APO. Curr Med Chem 16:3996-4002.Samuel CS, Hewitson TD. (2009) Relaxinand the progression of kidney disease.Curr Opin Nephrol Hypertens 18:9-14.Samuel CS, Mookerjee I, Halls ML,Summers RJ, Chew E, Bathgate RAD,Tregear GW, Hewitson TD. (2009)Investigations into the inhibitory effectsof relaxin on renal myofibroblastdifferentiation. Ann NY Acad Sci1160:294-299.Samuel CS, Royce SG, Chen B, Cao HF,Gossen JA, Tregear GW, Tang MLK. (2009)RXFP1 protects against airway fibrosisduring homeostasis, but not againstfibrosis associated with chronic allergicairways disease. Endocrinology 150:1495-1502.Scott DJ, Tregear GW, Bathgate RAD.(2009) Modeling the primary hormonebindingsite of RXFP1 and RXFP2. Ann NYAcad Sci. 1160:74-77.Sedaghat K, Finkelstein DI, GundlachAL. (2009) Effect of unilateral lesion ofthe nigrostriatal dopamine pathwayon survival and neurochemistry ofparafascicular nucleus neurons in therat-evaluation of time-course and LGR8expression. Brain Res 1271:83-94.Segura E, Albiston AL, Wicks IP, Chai SY,Villadangos JA. (2009) Different crosspresentationpathways in steady-stateand inflammatory dendritic cells. ProcNatl Acad Sci USA 106:20377-20381.Shabanpoor F, Hughes RA, BathgateRAD, Separovic F, Wade JD. (2009)Development of lanthanide-labeledhuman INSL3 as an alternative probe toradioactively labeled INSL3 for use inbioassays. Ann NY Acad Sci 1160:87-90.Shabanpoor F, Hughes RA, Zhang S,Bathgate RA, Layfield S, Hossain MA,Tregear GW, Separovic F, Wade JD. (2009)Effect of helix-promoting strategies onthe biological activity of novel analoguesof the B-chain of INSL3. Amino Acids38:121-131.Shabanpoor F, Separovic F, Wade JD.(2009) The human insulin superfamilyof polypeptide hormones. Vitam Horm80:1-31.Shemesh R, Hermesh C, Toporik A, LevineZ, Novik A, Wool A, Kliger Y, Rosenberg A,Bathgate RAD, Cohen Y. (2009) Activationof relaxin-related receptors by short,linear peptides derived from a collagencontainingprecursor. Ann NY Acad Sci1160:78-86.Smith CM, Lawrence AJ, SuttonSW, Gundlach AL. (2009) Behavioralphenotyping of mixed background(129S5:B6) relaxin-3 knockout mice. AnnNY Acad Sci 1160:236-241.Smith CM, Shen PJ, Ma S, Sutton SW,Gundlach AL. (2009) Verification of arelaxin-3 knockout/LacZ reporter mouseas a model of relaxin-3 deficiency. Ann NYAcad Sci 1160:259-260.Summers RJ, Bathgate RAD, Wade JD,van der Westhuizen ET, Halls ML. (2009)Roles of the receptor, the ligand, and thecell in the signal transduction pathwaysutilized by the relaxin family peptidereceptors 1-3. Ann NY Acad Sci 1160:99-104.Sutton SW, Shelton J, Smith CM, WilliamsJ, Yun S, Motley T, Kuei C, BonaventureP, Gundlach AL, Liu C, Lovenberg TM.(2009) Metabolic and neuroendocrineresponses to RXFP3 modulation in thecentral nervous system. Ann N Y Acad Sci.1160:242-249.Svendsen AM, Vrecl M, Knudsen L,Heding A, Wade JD, Bathgate RAD, DeMeyts P, Nøhr J. (2009) Dimerizationand negative cooperativity in the relaxinfamily peptide receptors. Ann NY AcadSci. 1160:54-59.Tang MLK, Samuel CS, Royce SG. (2009)Role of relaxin in the regulation of fibrosisin the lung: implications for airwayremodelling in asthma. Ann NY Acad Sci1160:342-347.Tregear GW, Bathgate RAD, Hossain MA,Lin F, Zhang S, Shabanpoor F, Scott D,Ma S, Gundlach AL, Samuel CS, WadeJD. (2009) Structure and activity in therelaxin family of peptides. Ann NY AcadSci 1160:5-10.van der Westhuizen ET, Wade JD, SextonPM, Summers RJ. (2009) Addition ofa carboxy-terminal green fluorescentprotein does not alter the binding andsignaling properties of relaxin familyPeptide receptor 3. Ann NY Acad Sci1160:105-107.Wade JD, Lin F, Hossain MA, ShabanpoorF, Zhang S, Tregear GW. (2009) Thechemical synthesis of relaxin and relatedpeptides. Ann NY Acad Sci 1160:11-15.Wraith DC, Pope R, Butzkueven H, HolderH, Vanderplank P, Lowrey P, Day MJ,Gundlach AL, Kilpatrick TJ, ScoldingN, Wynick D. (2009) A role for galanin inhuman and experimental inflammatorydemyelination. Proc Natl Acad Sci USA106:15466-15471.94

Neuro Research Services— Formerly Integrative Neuroscience FacilityOverviewNeuro Research Services (NRS) providesthree core technology platforms toFNI and the wider research community.These platforms encompass rodentneurohistology, advanced microscopy,and behavioural phenotyping. Eachfacility is maintained to the higheststandard and dedicated personnelprovide equitable access, trainingand technical guidance for users.FNI research staff and students havecontinued to access the facilities,equipment and services providedby the experienced NRS staff.The multi-user microscope facilitiesof the NRS provide advanced imagingcapabilities for researchers. Researchershave access to a suite encompassingtwo confocal imaging systems, A TIRF(Total Internal Reflection Fluorescence)microscope and several fluorescence,brightfield stereology microscopes andpost image analysis workstationsfor qualitative and quantitativeassessment of brain and othertissues. The neurohistology teamhave extensive knowledge of therodent brain morphology providingsupport to Florey researchersundertaking tissue preparation,staining, imaging and analysis.NRS has continued to provide Floreyresearchers with access to the mostcomprehensive range of rodentbehavioural testing facilities in Australia.As construction of the Melbourne BrainCentre (new Building) is well underway,NRS’s Travis Featherby was sponsoredto attend the Society for Neurosciencemeeting in 2009 to review the latesttechnology in rodent behaviour andwork undertaken by other prominentneurobehaviouralists, with a view toensure that we continue to provide andmaintain a state of the art platform forthe researchers moving into 2010-2011.Following the conference also he visitedthe newly commissioned Centre forMolecular Science, Vanderbilt Universityin Nashville, USA. He gained valuableinformation and made recommendationsthat will be implemented in the currentand new facilities.NRS continues to provide access to assistresearchers from other institutes anduniversities throughout Australia andoverseas, and offers contractresearch services to the commercialbiotechnology sector.Research ProjectsINF research projects for academiccollaboratorsBio21 InstituteCSIRO Food and Nutritional SciencesCSIRO Molecular & Health TechnologiesFlorey Neuroscience InstitutesInstitute for Molecular Bioscience,the University of QueenslandCell and Gene Therapy Group,Murdoch Children’s Research InstituteSkeletal Biology & Disease,Murdoch Children’s Research InstituteMitochondrial and Metabolic Research,Murdoch Children’s Research InstituteHormone Research,Murdoch Children’s Research InstituteQueensland Brain InstituteDepartment of Medicine,Royal Melbourne HospitalExternal academic access to INFequipment and resourcesMental Health Research InstituteMonash UniversityDepartment of Botany,University of MelbourneDepartment of Medicine,The University of MelbourneDepartment of Pathology,University of MelbourneDepartment of Pharmacology,University of MelbourneDepartment of Physiology,University of MelbourneFaculty of Veterinary Science,The University of MelbourneRMIT UniversityINF research projects for industryGlaxo Smith Kline (GSK)Achan Kham HoldingsFNI users of the INFNeural Plasticity GroupEmma BurrowsGrace ChanXin DuAnthony HannanMari KondoElizabeth ManningTerence PangMark RansomeJenni KarlThibault RenoirAddiction GroupCameron AdamsMichael BirdRobyn BrownBianca JuppElena KrstewAndrew LawrenceHeather MadsenMonique StagnittiNeuropeptides GroupJulia FreidgeimSherie MaCraig SmithPhilip RyanNilushiQian SangCedrick PeirsNeurochemistry LaboratoryPeta BurnsShanti DiwakarlaSandy (Seyoung) LeeLee Lee NgBroden Morgan96

Vi PhamManeesha WeerasingheHolly YeatmanNeurodegeneration LaboratoryTim AumannWah Chin BoonChris ByeHui Kheng ChuaBrette DyrekManal FargChathurini FernandoDeneice FongSuresh KaushikShi Sheng LuClare ParishJason SpencerLachlan ThompsonRogan TinsleyDoris TomasBrad TurnerAdam WalkerJerani PettikiriarachchiKylie WhiteIon Channels and Disease GroupElisa HillChris ReidKay RichardsEva SoVerena WimmerMolecular Neurobiology diseasesGroupHelena HyunLuning JiangShohreh MajdNeuropharmacology GroupLinda LauLinda MercerYea SeulRebecca SheeanNicole WallisMS Research GroupMelissa GresleAnna JonasGerry MaToby MersonLaura OluichJo StrattonAgnes WongStella ZhaoTraumatic Brain and Spinal CordInjury GroupJenny GunnersenJenny LakovicAlison MacintyreGoh Choo PengSynaptic Neurobiology LaboratoryLynley CordeiroDeniz KirikTomris MustafaMaria ShleperErnesto VargasSystems NeurophysiologySally HoodData generated in the INF wasincluded in a number of internationalpeer reviewed journal and conferencearticles.Scientific conferencesINF staff coordinated the FNI exhibitionat the IBRO Congress in Melbourne inJuly 2007, and attended the AusBiotechmeeting held in Brisbane in November2007 to meet with prospective clients.ManagementPlatform Management CommitteeDr Henry De Aizpurua, FNIProf Tony Bacic, Bio21 Institute,The University of MelbourneProf Malcolm Horne(Chair & Director NRS), FNIDr Kelvin Hopper, Innovation Dynamics,SydneyDr Ian Cooke, CNSBioMs Julie Anne Quinn(Secretary and Manager, NRS), FNIStaff ListIan Birchall B App Sc (RMIT) MSc (Melb) PhD(Swinburne)Mirjana Bogeski BSc (Hons) (Melb)Travis Featherby BSc (Hons) (Mon)Georgia Giannakis BSc (Hons) (Mon) PhD(Melb)Valérie Guille BSc MSc (Marseilles )PhD(Swinburne) (to 25/9/2009)Jacqueline Mills BSc (Hons) MAppSc (Syd)Brett Purcell Dip App Sc (Box Hill)Julie Anne Quinn B App Sc (RMIT) GDip Mktg(Mon) M Eng (RMIT)Angela Vais BSc (Hons) (Melb)97

Statistics andDecision SupportOverviewThe objective of the Statistics andInformatics Division is to provideexpertise in data, quantitative, andstatistical aspects of research projectscarried out by FNI. An important sourceof that expertise is our own researchin a number of areas of modellingmethodology of relevance for promotingthe use of high-standard, rigorousquantitative methods to support decisionmaking in the disciplines making upour institutional environment. Sinceadvanced statistical methodology is oflittle use for real applications without theavailability of appropriate computationaland modelling tools, adapting, extendingand validating complex statistical anddecision modelling software is anotherbasic task for our Division. The Divisionserves as a hub for collaboration withinFlorey Neuroscience Institutes andwith other Australian and internationalresearch institutions in the areas ofstatistical, data, and decision modelling.In 2009 Statistics and InformaticsDivision has provided statistical and datamanagement support to a number oflarge international clinical trials includingAVERTand EXTEND. In collaborationwith Neurosccience Trials Australia, theDivision has provided statistical supportfor SCIPA: a unique, multi-disciplinary,multi-centre program of research topromote neurological recovery, maintainhealth and wellness, and optimiseindependence following spinal cordinjury. In the area of Medical Informatics,the Division has lead the researchproject that successfully applied multiplecriteria decision analysis to chose themost appropriate computer softwarefor imaging the ischaemic penumbra inacute stroke patients. Novel applicationof case-based reasoning paradigm hassecured the success of the study offactors influencing specialists’ decisionsto mobilize patients after treatmentwith rtPA agent. In the area of healthservices research, in collaboration withCSIRO Mathematics and IS Divisionand The University of Melbourne, theDivision have built a process model ofrtPA delivery to acute ischaemic strokepatients in emergency departments.Effective decision support models forfast and appropriate rtPA delivery meansmore saved lives and less disability dueto more timely access to this cruciallyimportant treatment that can only beadministered within a relatively short timewindow. We further extend this work asa part of our on-going collaboration withSwinburne University of Technology withthe aim to create a simulation model forthe complete Stroke Chain of Survivaland Recovery. Such a decision supporttool that will allow to simulate variouscomplex scenarios resulting from theinteractions between prehospital, acute,and rehabilitation stroke care processes,thus providing the insights into the waysto better delivery of stroke care.Major Research ProjectsClinical decision support for acutestroke managementLeonid Churilov, Helen Deweyand Ka Lip ChewProcess-based risk analysis ofstroke careLeonid Churilov, Helen Dewey,Audrey Fridriksdottir and Andrew FlitmanComparative analysis ofepistemological assumptions in useof discrete-event simulation andsystems dynamics for health caresystems modellingLeonid Churilov, Sally Brailsford,Kristian Rotaru and Andrew FlitmanMulti-attribute evaluation of strokeimaging softwareLeonid Churilov, Geoff Donnanand Densheng LiuModelling of long-term iso-resourcegroups of stroke patientsLeonid Churilov and Helen DeweyUse of optimization algorithms formatched analisys of stroke trials dataLeonid Churilov and Adil BagirovModelling Factors and PreferencesInfluencing Clinical Decision Makingin Stroke Rehabilitation.Leonid Churilov, Julie Bernhardt,Jason Ha and Thomas LindenStaff and StudentsLeonid Churilov BsC(Hons), PhDLi Chun Quang Data ManagerTim Brewer IT Systems ManagerAudrey Fridriksdottir Research AssistantDensheng Liu Medical StudentConferences andPresentationsTowards Operational Risk – AwareInformation Systems: a Critical RealistPerspective at the 17th EuropeanConference on Information Systems,Verona, Italy (ECIS 2009).National and InternationalCollaborative LinksProf Andrew FlitmanSwinburne University,Melbourne, AustraliaDr Adil BagirovUniversity of Ballarat, AustraliaDr Kristian RotaruMonash University, AustraliaDr David SierCSIRO, AustraliaProf Sally BrailsfordSouthampton University, Southampton, UKProf Thomas LindedUniversity of Gothenburg, SwedenProf Marion RaunerUniversity of Vienna, AustriaEditorial PositionsA/Prof Leonid ChurilovMember, Editorial Board,Journal of Decision SystemsMember, Editorial Board,Intelligent Decision Technologies:International Journal98

PublicationsMONOGRAPHNeiger, D., Churilov, L., & Flitman, A.(2009). Value-Focused Business ProcessEngineering: a Systems Approachwith Applications to Human ResourceManagement. Springer. ISBN: 978-0-387-09520-2Journal PapersMa, H., Zavala, J.A., Teoh, H., Churilov,L., Gunawan, M., Ly, J., Wright, P., Phan,T., Arakawa, S., Davis, S.M., & Donnan,G.A. (2009). Fragmentation of theClassical Magnetic Resonance Mismatch“Penumbral” Pattern With Time. Stroke,40: 3752-3757.Ma, H., Zavala, J.A., Teoh, H., Churilov,L., Gunawan, M., Ly, J., Wright, P.,Phan, T., Arakawa, S., Davis, S.M., &Donnan, G.A. (2009). Penumbralmismatch is underestimated usingstandard volumetric methods and thisis exacerbated with time. Journal ofNeurology, Neurosurgery, and Psychiatry,80(9): 991-6.Sorbello, D., Dewey, H.M., Churilov,L., Thrift, A.G., Collier, J.M., Donnan,G.A., & Bernhardt, J. (2009). Very earlymobilisation and complications in the first3 months after stroke: Further results fromPhase II of A Very Early Rehabilitation Trial(AVERT). Cerebrovascular Diseases, Vol.28, Issue 4, pp.378-383.Sturmberg, J.P., Siew, E.-G., Churilov, L., &Smith-Miles, K. (2009). Identifying patternsin primary care consultations: a clusteranalysis. Journal of Evaluation in ClinicalPractice, Vol.15, No.3, pp.558-564.Neiger, D., Rotaru, K., & Churilov, L.(2009). Supply Chain Risk Identificationwith Value-Focused Process Engineering.Journal of Operations Management, Vol.27, No. 2, pp.154-168.Zhuang, Z.Y., Churilov, L., Burstein, F., &Sikaris, K. (2009). Combining data miningand case-based reasoning for intelligentdecision support for pathology orderingby general practitioners. EuropeanJournal of Operational Research 195(3):662-675.99

Stroke— Basic ScienceResearch Overview2009-10 has be en an exciting year forthe stroke basic science group. Highprofile papers such as the PLoS (SenaES, van der Worp HB, Bath PM, HowellsDW, Macleod MR. Publication bias inreports of animal stroke studies leadsto major overstatement of efficacy.PLoS Biol 2010;8(3):e1000344) publicationbias paper have had a major impactwith commentaries in the scientific(Nature, New Scientist) and lay (UK DailyTelegraph) press and our guidelinesfor avoiding bias at the bench havebeen adopted by the major journalsin our field. Experimental studies havealso led to significant publicationson stroke modelling and therapeuticstrategies including hypothermia. Anew collaboration with scientists fromCSIRO has facilitated a major strokebiomarkers project and clinical trial andthe recruitment of Professor HaraldSchmidt and his group from Monash hasallowed us to start investigations intothe damaging free radical mechanismsthat contribute to vascular and neuralinjury after stroke. Importantly, the mergerwith FNI has kick-started a range ofinternal collaborations including stemcell implantation and small moleculeneuroprotection studies with LachlanThompson, Clare Parish and Siew YinChai respectively.Major Research ProjectsCAMARADES: Systematic review andmeta-analysis of treatments for strokeDavid Howells, Malcolm Macleod,Emily Sena, Victoria O’Collins,Ulrich Dirngl and Philip BathThe CAMARADES collaboration forsystematic review and metanalysis of thedata supporting the efficacy of candidatetherapies for stroke (www.camarades.info)has continued to yield important results.Our CONSORT-like statement whichdetails how to avoid bias at the benchhas been adopted as Editorial Policy bythe journals Stroke, Journal of CerebralBlood Flow and Metabolism andInternational Journal of Stroke(1-3) andhas incorporated into the current STAIRguidlines(4). A particularly importantanalysis has been the demonstration thatpublication bias in stroke basic sciencecontributes to a >30% overestimation ofthe benefits of therapies trialed in thelaboratory(5). Journal of Cerebral BloodFlow and Metabolism will no acceptnegative but well conducted studies forpublication. This research program hasalso led to invitations to review the animalmodels used in the study of stroke(6), thevalue of animal experimentation as aprelude to clinical trials(7), and the areasof stroke biology most likely to lead tonew therapeutic options(8). Anothercritical outcome of the program has beenthe award of highly regarded Ph.D’s toDr Victoria O’Collins and Dr Emily Sena,we look forward to their heightenedinput at the post-doctoral level.Qualifiying more patients forthrombolysisDavid Howells, Victoria O’Collins, JennyFavaloro, Bill Wilson and Leeanne CareyThe EXTEND cinical trial of delayedthrombolytic therapy and biomarkerdiscovery has been funded by CSIRO($3 million) and the first patients recruited.The biomarker component of this largestudy aims to discover biomarkers ableto indicate where an individual is inthe temporal evolution of their stroke.This biological profile will provide abetter indication of chances of a goodoutcome and ability to respond safely tothrombolytic therapy than just the familiesrecollection of when the stroke occured.This simple concept will allow many morepatients to qualify for thrombolysis thanis currently possible with current timebasedtreatment restrictions. Experimentsin stroked animals where we can tightlycontrol our experimental conditionshave already revealed good candidatebiomarkers and confirmed that agenomic/proteomic approach will be lesswasteful of scarce resources than picking“fashionable” molecules for study.Mapping the long term plastic changesthat occur after strokeDavid Howells, Sarah Rewell, MichellePorritt, Emily Sena, Malcolm Mcleod,Mikael Jerndal, Kalle Forsberg, JenniferLees, Simon Koblar, Neil Spratt, HeideJenssen, Thomas Linden, Julie Bernhardtand Michael Nilsson.To be able to study long term plasticityafter stroke we have developed animalmodels of stroke in aged hypertensiveand diabetic animals that better reflect tothe aging co-morbid human populationaffected by stroke than the young healthyanimals normally used(9, 10) and haveperformed systematic reviews andmeta-analyses showing benefitof experimental approaches suchas environmental enrichment (11),Erythropoietin (12) and stem cell therapies(in press). Sarah Rewell has undertakenthe unprecedented task of strokesurgery on a large cohort of animals toenable us to map the behavioural andanatomical consequences of up to 6months survival after stroke. This workhas already shown which behaviouraldeficits are transient or persist long termand the immunohistochemical staining toallow us to map cellular, axonal, dendritic,and glial changes in relation to effectedvascular territories, neural connectivityand time have been performed and arebeing analysed.Hypothermia as a treatment forneural injuryDavid Howells, Peter Batchelor, MalcolmMacleod, Nicole Kerr, Amy Jeffrey’s,Emily Sena and Sarah RewellSystematic reviews and meta anlysis hasrevealed that hypothermia is one of themost potent neuroprotective strategiesavailable. Fully randomised and blindedexperiments have been performed toconfirm the results of our systematicreview and meta-analysis which pointedto hypothermia as one of the mostrobust neuroprotective strategies(13).In stroke models we have demonstratedthat the effect is robust but works bestwith smaller initial lesions and that usingpethidine to suppress shivering as wouldbe required in the clinic does not diminishthe effectiveness of hypothermia. Futureexperiments will determine whetherhypothermia enhances or reduces theeffectiveness of routine thrombolysisfor stroke and decompression for spinalcord injury. After spinal cord injury wehave demonstrated that hypothermiahas the potential to slow developmentof secondary spinal cord injury due topersistent compression and thus makedecompressive surgery a practicalproposition in the clinic(14). Clinicaltrials are being planned by the groupin both areas.100

Data mining to identify the bestcandidate stroke therapeuticsDavid Howells, Victoria O’Collins,Malcolm Macleod and Emily SenaWe are using systematic review and metaanalysis to probe our large database ofthe published data on the efficacy ofmore than 1000 drugs to determine whichremain good candidates after correctingfor experimental bias, lack of statisticalpower, and examination experimentalvariables which impinge on measuredoutcome. We have already ascertainedthat hypertension dramatically reducesthe efficacy of the majority of the smallsubset of drugs (>10% of the totaldataset) where this common comorbidityhas been considered. By comparing theexperimental profiles of therapy withtissue plasminogen activator in animalsand in man we have shown that whenmatched carefully, animal experimentaland clinical trial data give essentiallythe same data indicating that animalexperiments can be highly predictiveof human trials(15).The role of NOX genes in strokeHarald Schmidt, Emily Lam, Heide Ho,Liam Vo and David HowellsThere is an urgent need to move fromtreating symptoms of cardiovasculardiseases which include stroke, towardstreatment of the causes of disease. Aprerequisite for this is the identificationof the underlying mechanisms of disease.A major breakthrough in this endeavourhas been the realization that all knownrisk factors for heart attack and stroke(high blood pressure, diabetes, highcholesterol levels, smoking etc.) areassociated with ‘oxidative stress’, animbalance in the production andremoval of toxic molecules called freeradicals which attack the cells of bloodvessel walls leading to dysfunction andultimately disease. A major source offree radicals in the blood vessel wallis a family of enzymes called NADPHoxidases (NOX). Several studies haverecently demonstrated that upregulationNADPH oxidases may be involved in thedevelopment and progression of CVD,particularly stroke. We have recruited theSchmidt group to the stroke basic scienceteam to investigate the contributionof individual vascular NADPH oxidaseisoforms in the tissue damage andsymptoms observed post stroke and toexamine if NADPH oxidase inhibitionhas a protective effect. These studieswill include assessment of the roleof NOX4 in the development of dietinduced CVD using a unique geneticallymodified mouse strain, lacking theNADPH oxidase 4 isoform.In silico modelling of strokeDavid Howells, Victoria O’Collins,Fernando Calamante and Marc HommelSurvival of neural tissue after strokeis dependent on a complex series ofinteractiive processes ranging fromdegree or site of vessel occlussion,degree of vessel collateralisation, extendof underlying vessel damage and thecellular responses of the damaged orat risk tissues. These interactions aretoo complex to address effectivelyby serial animal experiments. Withcollaborations here in Melbourne, inthe USA and Europe we are developingan in silico model of the human androdent neurovascular systems thatwill permit rapid scenario testing andeffective focusing of animal and humanexperiments. A similar modellingapproach is being used to identify thekey points of the ischaemic cascadeto develop a set of common surrogateoutcome markers that can be usedidentically in both animals and man isthe part of a current CRC application.Other Research ProjectsAmelioration of macrophagecytotoxicity after spinal cord injury:impact on secondary injury andaxonal regenerationBatchelor, Loy, HowellsRegeneration of the corticospinaltract in the injured spinal cordBachelor, Wills, HowellsEffects of interleukin- 4 on macrophageand microglia activation after spinalcord injuryBachelor, Kerr, Sidon, Favaloro, HowellsBiological evaluation of novelradioligands for the imagingof hypoxic tissue in stroke (15)Howells, Ackermann, SprattDetermination of cerebral bloodflow in the ischaemic penumbraHowells, Spratt, O’CollinsThe Therapeutic Potential ofNeuroepithelial Cells in the InjuredRat Spinal CordBatchelor, WillsMacrophage phenotypes and theireffect in the Central Nervous SystemLoy, Batchelor, HowellsNHMRC PROGRAM: Developingnew therapeutic targets for strokeDonnan, Howells, Hankey, Calford, DavisSYSTEMS BIOLOGY: Human Stemcells as a test bed for evaluatingstroke therapeuticsHowells, AntonicTissue plasminogen activator (tPA)in Parkinson’s DiseaseHowells, Liberatore, MedcalfStaff and StudentsDavid Howells BSc, PhDPeter Batchelor MBBS, FRACP, PhDJenny Favoloro PhDMichelle Porritt PhDKate Sidon BSc (Med Sci) (Hons)Victoria O’Collins PhD Bsci, BA(Hons) LLB,BcomSarah Rewell BSc(Hons)Candace Loy BSc (Hon), PhD CandidateTaryn Wills BSc (Hons), PhD CandidateAnna Antonic MSc (Hon)Emily Lam BSc, PhDHeide Ho BScLiam Vo BSc101

Stroke— Basic ScienceMajor Collaborative LinksDr Malcolm MacleodUniversity of Edinburgh, UKProf Nancy RothwellManchester, UKDr Michael NilssonGotteborg, SwedenProf Marc HommelGrenoble, FranceProf John MorserStanford, USAProf Robert MedcalfCentre for Blood Diseases,Monash UniversityProf Harald SchmidtUniversiteit Maastricht, The NetherlandsDr Spratt and Dr ParsonsUniversity of Newcastle, NSW.Editorial PositionsA/Prof David HowellsThe International Journal of StrokeExperimental & Translational StrokeMedicineTranslational Stroke ResearchVisiting ScientistsDr Emily SenaDr Malcolm McLeodProf Harald SchmidtProf Michael NilssonPublicationsMacleod MR, Fisher M, O’Collins V, SenaES, Dirnagl U, Bath PM, et al. Reprint:Good laboratory practice: preventingintroduction of bias at the bench. Int JStroke. 2009 Feb;4(1):3-5.Macleod MR, Fisher M, O’Collins V, SenaES, Dirnagl U, Bath PM, et al. Reprint:Good laboratory practice: preventingintroduction of bias at the bench. J CerebBlood Flow Metab. 2009 Feb;29(2):221-3.Macleod MR, Fisher M, O’CollinsV, Sena ES, Dirnagl U, Bath PM, et al.Good laboratory practice: preventingintroduction of bias at the bench. Stroke.2009 Mar;40(3):e50-2.Fisher M, Feuerstein G, Howells DW,Hurn PD, Kent TA, Savitz SI, et al. Updateof the stroke therapy academic industryroundtable preclinical recommendations.Stroke. 2009 Jun;40(6):2244-50.Sena ES, van der Worp HB, Bath PMW,Howells DW, Macleod MR. PublicationBias in Reports of Animal Stroke StudiesLeads to Major Overstatement of Efficacy.PLoS Biol. 2010;8(3):e1000344.Howells DW, Porritt MJ, Rewell SS,O’Collins V, Sena ES, van der Worp HB, etal. Different strokes for different folks: therich diversity of animal models of focalcerebral ischemia. J Cereb Blood FlowMetab. 2010 May 19.van der Worp HB, Howells DW, SenaES, Porritt MJ, Rewell S, O’Collins V, etal. Can animal models of disease reliablyinform human studies? PLoS Med.2010;7(3):e1000245.Howells DW, Donnan GA. Wherewill the next generation of stroketreatments come from? PLoS Med.2010;7(3):e1000224.Rewell SS, Fernandez JA, Cox SF, SprattNJ, Hogan L, Aleksoska E, et al. Inducingstroke in aged, hypertensive, diabetic rats.J Cereb Blood Flow Metab. 2010 Jan 13.Porritt MJ, Chen M, Rewell SS, Dean RG,Burrell LM, Howells DW. ACE inhibitionreduces infarction in normotensive butnot hypertensive rats: correlation withcortical ACE activity. J Cereb Blood FlowMetab. 2010 Apr 21.Janssen H, Bernhardt J, Collier JM, SenaES, McElduff P, Attia J, et al. An enrichedenvironment improves sensorimotorfunction post ischemic stroke.Neurorehabilitation and Neural Repair.2010;In press.Jerndal M, Forsberg K, Sena ES, MacleodMR, O’Collins VE, Linden T, et al. Asystematic review and meta-analysis oferythropoietin in experimental stroke.J Cereb Blood Flow Metab. 2010May;30(5):961-8.van der Worp HB, Sena ES, Donnan GA,Howells DW, Macleod MR. Hypothermiain animal models of acute ischaemicstroke: a systematic review and metaanalysis.Brain. 2007 Dec;130(Pt 12):3063-74.Batchelor PE, Kerr N, Gatt AM, AleksoskaE, Cox SF, Ghasem-Zadeh A, et al.Hypothermia prior to decompression:Buying time for treatment of acute spinalcord injury. Journal of neurotrauma. 2010May 26.Sena ES, Briscoe CL, Howells DW,Donnan GA, Sandercock PA, Macleod MR.Factors affecting the apparent efficacyand safety of tissue plasminogen activatorin thrombotic occlusion models of stroke:systematic review and meta-analysis. JCereb Blood Flow Metab. 2010 Jul 21.102

Stroke— Epidemiology and Public HealthOverview: EpidemiologyThe main aims of our research activitiesin the Epidemiology Division are todetermine how stroke impacts on patientsand Australian society. Over the last 13years, in collaboration with A/ProfessorAmanda Thrift at the Baker IDI, Heart andDiabetes Institute we have been focussedon the North East Melbourne StrokeIncidence Study (NEMESIS) which initiallyenrolled stroke patients in 1996-98. Agroup of more than 1600 stroke patientswere enrolled and followed in this studyto assess how they fared in the long term.The 10 year follow ups for the final year ofthe cohort were completed in 2009.Information collected from NEMESISover 10 years of follow-up, has allowedus to update our estimates of the costsand burden of stroke in Australia. Wenow estimate that the total lifetimecosts for all cases of first ischaemic andhaemorrhagic stroke that occurred in2004 are approximately $2 billion. Duringthe first year after stroke, haemorrhagicstrokes are more expensive to treatthan ischaemic strokes. However, overa lifetime, ischaemic strokes are morecostly per case than haemorrhagicstrokes ($64,733 versus $54,721). Overall,outpatient and community costs aregreater than the costs of inpatienthospital care. These data will be useful forplanning health service requirements tomeet the growing needs of society. This isbecause there is strong evidence that thenumber of stroke events will increase asthe population ages.Other projects in the Epidemiologydivision have included BAT24, a studyof BP patterns and outcomes in patientswith TIA, compared to age and sexmatched controls.Overview: Public HealthThe research activities of the PublicHealth Division for 2009 have includeda range of projects related to improvingclinical management of stroke anddisease prevention. Research quantifyingthe quality of stroke care in publichospitals continues to be an importantarea of research. Examples include theestablishment of the first Australianclinical quality registry for stroke andinvolvement in various national auditand evaluation programs, as well as morespecific assessments lead by our groupin Victoria and New South Wales. Wehave continued to work with the NationalStroke Foundation on several publichealth programs (eg Know Your Numbersblood pressure program) and healthservices evaluations (eg national auditof post-acute stroke services) providingacademic and statistical contributions.This year we are also collaborating withthe Victorian Department of Health,Bendigo Health, the Loddon MalleeRural health Alliance, Ambulance Victoriaand the National Stroke Foundation onan exciting new project examining theeffectiveness of a stroke Telemedicineprotocol to improve the use ofintravenous thrombolysis in regional areas.Major Research ProjectsThe Victorian StrokeTelemedicine ProjectChris Bladin, Dominique Cadilhac, SoniaDenisenko, Helen Dewey, Peter Disler,Tony Walker, Bruce Winzar and Ian MosleyThe overall objectives of this project areto investigate the potential benefits andvalue of telemedicine for enhancingcapacity in rural and regional hospitalsto provide increased access to acutestroke treatments. The Victorian StrokeTelemedicine (VST) program has beendesigned to demonstrate the use of aformalised telemedicine protocol in aregional Victorian health service(Bendigo Health). The VST programincludes access to a pool of external(Melbourne-based) physicians withexpertise in stroke using videoconferencingtechnology (telemedicine).Clinical decision-making is enhanced byhaving the ability to perform joint clinicalconsultations with the view to buildingsustainable, long term capacity andcapability. The program also includesuse of community, ambulance staff andclinician education to raise awarenessand knowledge about stroke. This projecthas been co-funded with a VictorianScience Investment Fund Departmentof Innovation, Industry and RegionalDevelopment (DIIRD) grant.Australian Stroke Clinical Registry(AuSCR)Dominique Cadilhac, Natasha Lannin,Craig Anderson, Amanda Thrift, ChrisPrice and Geoffrey DonnanIn August 2008, the AustralianCommission on Safety and Quality inHealth Care released a Request ForTender to test and validate the draft‘Operating Principles and TechnicalStandards for Australian Clinical QualityRegistries. A consortium was formedbetween the NSRI, The George Institutefor International Health, the NationalStroke Foundation and the Stroke Societyof Australasia who submited a successfulproposal for an Australian Stroke ClinicalRegistry (AuSCR).The main aim of the Registry is to ensurea systematic way of monitoring andproviding evidence for improvementsin the quality of stroke care throughoutAustralia. The Registry includes aminimum data-spine and is not intendedto be a comprehensive data collectiontool. Principal features include limitingthe amount of manual data entry;achieving 100% case ascertainment; andconducting a 3-month patient followupquestionnaire. An experiencedTechnology company has been appointedto facilitate harmonisation of the Registrywith mainframe hospital systems usingstrict security processes and systems.The Registry is governed by a nationallyrepresentative Steering Committeechaired by Professor Sandy Middleton.The pilot must be completed byNovember, 2009.North East Melbourne StrokeIncidence Study (NEMESIS)Geoff Donnan, Amanda Thrift, HelenDewey, Richard Macdonell and Rob CarterThis study was designed to determine theincidence and outcome of stroke and itssubtypes in north east Melbourne. Thestudy commenced in 1996 and 10 yearfollow ups were completed in 2009.103

Stroke— Epidemiology and Public HealthOther Research ProjectsEvaluation of the Victorian StrokeCare StrategyThe aim of this evaluation is to describecurrent clinical practice for stroke asdescribed by health professionals workingwithin ten area health services. Thesedata will provide evidence to support thedevelopment of implementation plansand will be used as a baseline descriptionof hospital services in future assessmentsof the impact of the Victorian StrokeCare Strategy.New South Wales Health StrokeProgram EvaluationTime series clinical audits of hospitals inboth metropolitan and rural NSW since2003. Aggregated data have been usedto provide evidence that stroke careimprovement initiatives have significantlyreducing disability following stroke.Differences in Stroke Unit Care:Melbourne versus Trondheim (Norway)This Master of Physiotherapy project wasdesigned to provide evidence of thedifferences in processes of care betweenstroke units at Austin Health and St OlavsHospital in Trondheim, Norway (GoldStandard Care). The thesis was submittedin December 2009.A Phase II multi centred, Single Blind,Randomised, Controlled Trial of theStroke Self Management Program(SSMP) versus Standard Care or theChronic Condition Self ManagementProgram in Stroke SurvivorsMalcolm Battersby, Sally Hoffman,Dominique Cadilhac, Richard Lindley,Richard Osborne and Erin LalorThe aim of this Phase II randomizedcontrolled trial (RCT) was to evaluatewhether a chronic disease selfmanagementprogram designedspecifically for patients with stroke wassafe and feasible compared to standardcare or a generic program. A total of 143participants were randomised across thestudy into two intervention groups orstandard care. Final follow-up of patientsoccurred in 2009.BAT 24 – Blood pressure in Ambulatorymonitoring after Transient ischemicattack (TIA) or minor strokeTransient Ischaemic Attack (TIA) is awell-recognized risk factor for stroke andhypertension is a major risk factor forTIA and stroke. Furthermore, differentBP patterns have been described usingAmbulatory Blood Pressure Monitoring(ABPM) and a “non-dipping” pattern iean absence of the normal reduction innocturnal BP is shown to be associatedwith stroke and a worse prognosisfor target organ damage. On thebackground of insufficient data about thecommon BP pattern after TIA and minorstroke and the relationship betweenthe different BP patterns and TIA/minorstroke outcomes, this natural history casecontrol study is being conducted.The purpose of this project is to providenew information about the associationbetween BP variability and, autonomicnervous system function in TIA or minorstroke patients and the subsequent risksof recurrent TIA or stroke by assessingHeart Rate Variability and 24-hour ABPMfor TIA/minor stroke patients within 7days after symptoms onset as well asfor control participants and assessingoutcomes at 3 months.Additional Research ProjectsKnow Your Numbers programevaluationDominique Cadilhac, Monique Kilkennyand Roslyn JohnsonA Phase II multi centred, Single Blind,Randomised, Controlled Trial of theStroke Self Management Program(SSMP) versus Standard Care or theChronic Condition Self ManagementProgram in Stroke SurvivorsMalcolm Battersby, Sally Hoffman,Dominique Cadilhac, Richard Lindley,Richard Osborne and Erin LalorNational Stroke Services Audit:acute and rehabilitation servicesDominique Cadilhac, Monique Kilkenny,Erin Lalor, D Harris and Graeme HankeyThe Economic Benefits ofReducing Disease Risk FactorsDominique Cadilhac, Anne Magnus, DoraPearce, Lauren Sheppard, Toby Cumming,Theo Vos and Rob CarterNew South Wales stroke audit programDominique Cadilhac, Monique Kilkenny,Geoff Donnan and Chris LeviStand FirmDominique Cadilhac, Amanda Thrift,Richard Gerraty, Thanh G. Phan, ChrisBladin, SM Fitzgerald and G TikellisQuality in Acute Stroke Care ProjectDominique Cadilhac, Sandy Middleton,Chris Levi, J Ward and R GriffithsICARUSSDominique Cadilhac, Helen Dewey,Geoff Donnan and Jacques JoubertNSW Rural Stroke ProjectDominique Cadilhac, Chris Levi, MPollack, M Gill, L Cutler and M LongworthAbsolute Risk Assessment GuidelinesProjectDominique Cadilhac, T Tonkin,S Colagiuri, T Matthew and Erin LalorVictorian Stroke Care StrategyEvaluationDominique CadilhacNew Zealand Cost of Illness StudyDominique CadilhacBAT 24Wenwen Zhang, Dominique Cadilhac,Geoff Donnan and Helen DeweyStroke Units Trondheim and MelbourneTara Purvis, Dominique Cadilhac,Geoff Donnan and Julie Bernhardt104

NEMESIS: Long-term outcome afterstroke: Survival, stroke recurrence,functional ability and costsHelen Dewey, Richard Macdonelland Mandy ThriftStaff and StudentsHelen M. Dewey MB BS, PhD, FRACP,FAFRM (RACP)Sue Mosley Hon Research NurseMary Staios Hon Research NurseWenwen Zhang MDDominique Cadilhac PhD, MPubHlth,BNurs (post-reg), RNMonique Kilkenny B Appl Sci (MRA),G Dip (Epidemiol & Biostats), MPH (Research)Natasha Lannin PhD (honorary appointment)Ian Mosley MB (Management), GradDip(Health Admin), Grad Cert (education), GradDip(Education), BB (Accounting), RN (honoraryappointment)Karen Moss BBSc (Hons)Conferences andPresentationsStroke Society of Australasia,Cairns Platform 9/09 (Cadilhac)Stroke Society of Australasia, CairnsInvited Workshop 9/09 (Cadilhac/ Lannin)Shoalhaven Division of GeneralPractice Nowa (6/5) and Milton Ulladulla(7/5) (Cadilhac)Florey Postdoctoral Association annualscientific meeting (10/09) (Cadilhac)Australian Commission on Safety andQuality in health care: pilot registriesproject workshop, Melbourne (02/09)(Cadilhac)National and InternationalCollaborative LinksProf Craig AndersonGeorge Institute, AustraliaAuSCRDr Natasha LaninUniversity of NSW RehabilitationStudies Unit, AustraliaAuSCRA/Prof Amanda ThriftBaker IDI, AustraliaNEMESIS, AuSCR and STAND FIRM forstroke patients (RCT)A/Prof Theo VosUniversity of Queensland, AustraliaThe economic benefits of reducingdisease risk factorsProf Robert CarterDeakin University, AustraliaThe economic benefits of reducingdisease risk factorsMs Anne MagnusDeakin University, AustraliaThe economic benefits of reducingdisease risk factorsA/Prof Chris LeviHunter Stroke Service, NSW AustraliaNSW Health Stroke Program EvaluationMr Mark LongworthNew South Wales Health, AustraliaNSW Health Stroke Program EvaluationDr Erin LalorNational Stroke Foundation, AustraliaNSF National Audit Program. Phase II,multicentred, single blind randomisedcontrolled trial of the “Stroke selfmanagementprogram’ versusstandard care. National BloodPressure Awareness Program.Prof Malcolm BattersbyFlinders University, AustraliaPhase II, multicentred, single blindrandomised controlled trial of the“Stroke self-management program’versus standard care.Prof Sandy MiddletonAustralian Catholic University, AustraliaFever, hyperglycaemia and swallowingdysfunction management in acute stroke:A cluster randomised controlled trial ofknowledge transfer & AuSCR.Editorial/Advisory BoardPositionsDr Dominique CadilhacClinical Audit (Dove Medical Press):honorary editorial boardStroke Society of Australasia executivecommitteeNational Advisory Committee: NationalStroke Audit of Hospitals, National StrokeFoundationNational Vascular Disease PreventionAllianceStroke Guidelines Advisory Committee:National Stroke FoundationA/Prof Helen DeweyInternational Journal of StrokeStrokeBoehringer Ingelheim Stroke AdvisoryBoard (Australia)AwardsDominique Cadilhac:National Heart Foundation Postdoctoralpublic health training fellowship(cofunded NHMRC)Finalist VicHealth Awards category:Knowledge and Understanding for“Economic Benefits of Prevention”.Melbourne School of Population HealthResearch: Head’s Award for DoctoralExcellenceNational Heart Foundation Travelgrant ($2,000)105

Stroke— Epidemiology and Public HealthPublicationsArticlesCrosbie DC, Hariharan S, SimpsonMA, Walker SP, Dewey HM, Reade MC.Late onset ornithine transcarbamylasedeficiency: a potentially fatal yet treatablecause of coma. Critical Care andResuscitation 2009;11:222-227.Zhang WW, Dewey H, Cadilhac DA,Donnan G. Emerging Risk Factors ofStroke. Medicine Today 2009;10(11):35-41.Sorbello, D., Dewey, H. M., Churilov, L.,Thrift, A. G., Collier, J. M., Donnan, G.,Bernhardt, J. Very early mobilisation andcomplications in the first 3 months afterstroke: Further results from Phase II ofA Very Early Rehabilitation Trial (AVERT).Cerebrovascular Diseases 2009;28:378-383Price C, Blacker D, Grimley R, Dewey H,Gerraty R, Koblar S, Denisenko S, StoreyC, Bladin C, Hill K. National survey ofmanagement of transient ischaemicattack in Australia: Take ImmediateAction. The Medical Journal of Australia2009;191:17-20.Zhang W. W, Cadilhac D. A, DonnanG. A, O’Callaghan C and Dewey H. M.Hypertension and TIA. InternationalJournal of Stroke 2009;4:206-214.Impact Factor: 1.917Cadilhac DA, Carter R, Thrift AG andDewey HM. Estimating the long-termcosts of ischemic (IS) and hemorrhagic(ICH) stroke for Australia: new evidencederived from the North East MelbourneStroke Incidence Study (NEMESIS). Stroke2009;40:915-921. Impact Factor: 6.296.Gall SL, Dewey HM and Thrift AG.Smoking cessation at 5 years afterstroke in the North East MelbourneStroke Incidence Study (NEMESIS).Neuroepidemiology 2009;32:196-200.Iwanaga T, Arakawa S, Siritho S, Fitt G,Dewey HM, Chambers BR and DonnanGA. Paracentral strip infarcts of themiddle cerebral artery: borderzoneischemia or cortical artery occlusion?Cerebrovascular Diseases; 2009;27:215-222.Thrift, AG, Dewey, HM, Sturm, JW,Srikanth, VK, Gilligan, AK, Gall, SL,Macdonell, RAL, McNeil, JJ and Donnan,GA. Incidence of Stroke Subtypes in theNorth East Melbourne Stroke IncidenceStudy (NEMESIS): Differences betweenMen and Women. Neuroepidemiology,32:11-18, 2009. Times Cited: 1Nazir, FS, Petre, I and Dewey, HM.Introduction of an Acute Stroke Team:An effective approach to hastenassessment and management of strokein the emergency department. J ClinNeurosci, 16:21-5, 2009.Battersby M, Hoffmann S, Cadilhac D,Osborne R, Lalor E, Lindley R. ‘Gettingyour life back on track after stroke’: aPhase II multi-centered, single-blind,randomized, controlled trial of theStroke Self-Management Program vs.the Stanford Chronic Condition Self-Management Program or standard carein stroke survivors. Int J Stroke2009;4(2):137-44.Cadilhac DA, Moodie ML, Lalor EE.DALYs and Public Health Programs forStroke: Australian perspectives. In: PreedyVR, Watson RR, editors. Handbook ofDisease Burdens and Quality of LifeMeasures. September ed. New York:Springer, 2009:4448.Cadilhac DA, Moss K. Stroke care inselected Victorian health services: PhaseI Evaluation Report. Victorian Stroke CareStrategy. Heidelberg Heights: NationalStroke Research Institute, 2009:84.Cadilhac DA, Magnus A, Cumming TB,Sheppard L, Pearce D, Carter R. TheEconomic Benefits of Reducing DiseaseRisk Factors: Summary Report. preparedfor VicHealth. Burwood: Deakin Universityand the National Stroke ResearchInstitute, 2009:65.Cadilhac DA, Magnus A, Cumming TB,Sheppard L, Pearce D, Carter R. TheEconomic Benefits of Reducing DiseaseRisk Factors: Research Report. preparedfor VicHealth. Burwood: Deakin Universityand the National Stroke ResearchInstitute, 2009:291.Cadilhac DA, Lannin N, Anderson C, LimJ, Middleton S, Faux S, et al. Testingand Validation of the Draft OperatingPrinciples and Technical Standards forAustralian Clinical Quality Registries:Pilot Project Final Report AustralianStroke Clinical Registry (AuSCR) version1.1. Heidelberg Heights: National StrokeResearch Institute, 2009:95.Harris D, Cadilhac D, Kilkenny M,Read S, Boddice G, Ritchie E, et al.National Stroke Audit Acute ServicesOrganisational Survey Report 2009:National Stroke Foundation, 2009:51.Harris D, Ritchie E, Cadilhac D, KilkennyM, Read S, Boddice G, et al. NationalStroke Audit Acute Services Clinical AuditReport 2009. Melbourne: National StrokeFoundation, 2009:44.Joubert J, Joubert LB, de Bustos EM,Ware D, Jackson D, Harrison T, et al.Telestroke in stroke survivors. CerebrovascDis 2009;27 Suppl 4:28-35.Kilkenny MF, Johnson R, Cadilhac DA.Feasibility of a pilot program to increaseawareness of blood pressure as animportant risk factor for stroke in Australia.Int J Stroke 2009;In press 19 October.Levi CR, Lindley R, Smith B, BladinC, Parsons M, Read S, et al. Theimplementation of intravenous tissueplasminogen activator in acute ischaemicstroke - a scientific position statementfrom the National Stroke Foundation andthe Stroke Society of Australasia. InternMed J 2009;39(5):317-24.Middleton S, Levi CR, Ward J, GrimshawJ, Griffiths R, D’Este C, et al. Fever,hyperglycaemia and swallowingdysfunction management in acute stroke:A cluster randomised controlled trial ofknowledge transfer. ImplementationScience 2009;4(16):Open Access.Purvis T, Cadilhac DA, Donnan GA,Bernhardt J. Systematic review of processindicators: including early rehabilitationinterventions used to measure qualityof acute stroke care. Int J Stroke2009;4(2):72-80.Purvis T. Determining differences inStroke Uni Care: Melbourne versusTrondheim. Master of PhysiotherapyThesis, The University of Melbourne,December 2009: 254.106

Conference PapersDewey HM, Lees J, Payne K, ChurilovL. Place of residence and patterns ofresource use long term after stroke arepredicted by disability: Australian findingsfrom the stroke patients resource use andcaregiver burden outcomes by severitystudy (recovery). International Journalof Stroke 2009;4(Suppl. 1):23.Mosley I, Nicol M, Donnan G, Dewey H. Aretrospective assessment of acute strokepatients in the prehospital setting usingthe fast criteria. International Journal ofStroke 2009;4(Suppl. 1):14.Simpson M, Dewey H, Churilov L, BladinC, Markus R, Ahmed N, Parsons M.Thrombolysis for acute stroke in Australia:experience from the SITS-InternationalStroke Thrombolysis Register 2002-2008. International Journal of Stroke2009;4(Suppl. 1):13.Mosley I, Nicol M, Donnan G, DeweyH. Stroke knowledge among membersof the general public. How much isenough? International Journal of Stroke2009;11(Suppl. 1):11.H Dewey, J Collier, D Leys, J BernhardtLittle physical activity in a French strokeunit: results from an observational study.Poster presentation, 18th EuropeanStroke Conference, Stockholm, Sweden,May 26-29, 2009. CerebrovascularDiseases 2009;27(suppl 6):179.Simpson, Marion; Crosbie, David; Reade,Michael; Walker, Sue; Cousins Alison;Dewey, Helen. A 39-year old womanwith coma in early pregnancy. Posterpresentation, Annual Scientific Meetingof the Australian and New ZealandAssociation of Neurologists, Christchurch,New Zealand, 18-21 May 2009.Cadilhac DA Lannin N, Kilkenny M,Anderson C, Thrift AG, Price C, et al.Establishment of the new AustralianClinical Stroke Registry (AuSCR) minimumdataset. [ABSTRACT 2B.5]. Int J Stroke2009;4 (Suppl.1):12.Cadilhac DA, Lannin N, Kilkenny M,Anderson C, Price C, Thrift AG, et al.Establishment of the minimum dataset forthe new Australian clinical stroke registry(AuSCR). [ABSTRACT 9 Management& Economics]. Cerebrovasc Dis2009;27(Suppl 6):235.Middleton S, Levi CR, D’Este C, Dale S,Griffiths R, Grimshaw J, et al. Ninety-daymortality and morbidity postacute stroke:baseline results from the quality in acutestroke care project. [ABSTRACT 2A.1].Int J Stroke 2009;4 (Suppl.1):9.Harris D, Cadilhac DA, Faux S, GillisC, Hubbard I, Kilkenny M, et al. Strokerehabilitation in Australia: audit 2008results. [ABSTRACT A8]. Int J Stroke2009;4 (Suppl.1):21.Lannin N, Cadilhac D, Anderson C, LeviCR, Faux S, Price C, et al. Challengesto the establishment of the Australianstroke clinical registry to promote qualityimprovement in stroke care. [ABSTRACTA3]. Int J Stroke 2009;4 (Suppl.1):20.107

Stroke— Stroke Imaging and UltrasoundImaging – OverviewMajor Research ProjectsThe ischaemic penumbra continues tobe an important focus of our research.This is brain tissue which, while damaged,continues to live after the onset of thestroke process. The duration of this isuncertain in humans although we nowhave the ability to create images ofits behaviour using positron emissiontomography (PET) and magneticresonance imaging (MRI).Using MR we have shown thatsalvageable (penumbral) brain tissue maylast up to 48 hours post stroke onset andthat the distribution of this salvageabletissue becomes more variable withtime. Further, improved computer coregistrationtechniques may identifymore patients than previously realisedwho have penumbral tissue. This mayexpand the number of patients eligiblefor treatment with thrombolytic agentssuch as tPA.Another intiative of significance is the useof PET using 18F-PIB to image amyloidplaque in patients with recent onsetischaemic and haemorrhagic stroke.There are interesting links betweenvascular dementia, stroke and Alzheimer’sdisease which may be, in part, unravelledusing this technique.Ultrasound – OverviewWe are completing an investigationconcerning the clinical significance ofa newly appreciated ultrasound sign,referred to as “small vessel knock”. TheNSRI is collaborating with CompumedicsDWL, a Melbourne-based transcranialDoppler ultrasound company, todetermine whether knock is useful inthe diagnosis of stroke. Dr ChesdaUdommongkol, a stroke researchfellow from Bangkok, is completing hisPhD study of knock, due for completionin 2010.Dr Udommongkol’s work has found thatthere are other causes of knock-likesignals including wall-motion artefact. Incollaboration with CSIRO, and a groupin Leicester UK, he created a bench-topscaled-up model of a vessel bifurcationperfused by blood-mimicking fluid and apump simulating normal pulatile flow.In vivo 11C-PiB binding is increasedin patients with Cerebral AmyloidAngiopathy related Haemorrhage.John V Ly, Geoffrey A Donnan, Victor LVillemagne, Jorge A Zavala, Henry Ma,Graeme O’Keefe, Sylvia J Gong,Rico M Gunawan, Tim Saunder,Uwe Ackerman, Henri Tochon-Danguy,and Christopher C RoweThe in vivo-diagnosis of cerebral amyloidangiopathy (CAA) is inferred from clinicaland structural imaging features. 11C-PiBis a PET ligand that binds to beta-amyloid(Ab) both in extracellular plaques andvessel walls. We hypothesized thatpatients with a clinical diagnosis of CAArelated ICH (CAAH) have increased11C-PiB uptake and that the patterndiffers from Alzheimer’s disease (AD).Patients with CAAH based on establishedclinical criteria were studied using 11CPiB PET and compared to age matchedcontrols and AD patients. DistributionVolume Ratio (DVR) parametric maps werecreated using the cerebellar cortex as areference region. 11C-PiB binding wasmoderately increased in most patientswith probable CAAH. The distributionmay differ from that seen in AD. 11C-PiBPET may assist the in-vivo diagnosis ofCAA and serve as a surrogate marker forfuture therapeutic studies.Figure 1Figure 1: Representative axial DVR PiB PETimages of a normal control subject and patientswith Cerebral Amyloid Angiopathy Haemorrhage(CAAH) (with corresponding CT showinghaemorrhage in right parieto-occipital region) andAlzheimer’s Disease (AD).Figure 2: Representative PiB PET image andcorresponding CT scan of thrombolysis (tPA)treated ischemic stroke patients with (toppanel) and without (bottom panel) ParenchymalHemorrhage (PH).108

CAA detected by PiB PET predisposesto tPA related haemorrhageJohn V Ly, Christopher C Rowe, Victor LVillemagne, Jorge A Zavala, Henry Ma,Graeme O’Keefe, Sylvia J Gong, RicoGunawan, Leonid Churilov,Tim Saunder,John Sachinidis, Uwe Ackerman, HenriTochon-Danguy and Geoffrey A DonnanIncreasing evidence suggests thatCerebral Amyloid Angiopathy (CAA)may be an important predisposingfactor for the haemorrhagiccomplications of rtPA therapy. N-methyl-[11C]2-(4’-methylaminophenyl)-6-hydroxybenzothiazole (11C PiB) is aPET amyloid ligand which also binds tobeta-amyloid in cerebrovascular walls.We hypothesized that patients whodeveloped parenchymal haemorrhage(PH) after tPA may have increased 11CPiB PET retention compared to thosewith no haemorrhagic complications.Patients treated within 3 hours of onsetof ischaemic stroke with tPA were studiedusing PET to compare PiB retention inthose with and without parenchymalhaematoma formation and normal agematched controls. Ischaemic strokepatients treated with rtPA who developPH were shown to have higher neocorticalPiB retention compared to those withoutPH, suggesting underlying CAA as apredisposing factor for tPA relatedhaemorrhage. The high sensitivity andspecificity for this finding may providean impetus for the development of amore practical rapid pre treatmentscreening technique.Figure 2Other Research ProjectsPreconditioning stroke fMRIGeoff Donnan and Amy BrodtmannThe human brain is a remarkably plasticorgan, capable of reorganisationfollowing injury even until the ninthdecade. This reorganisation – orremodelling – has been well describedfollowing stroke, but the drivers anddeterminants of this phenomenon remainpoorly understood. It is understoodthat a lack of blood supply, such as inischaemic stroke, leads to utilisation ofother areas of the brain, both adjacentto the injury and in other distant butfunctionally connected cortex. However,it is not known whether reduced bloodsupply, such as occurs in patients witharterial narrowing (stenoses), resultingin chronic hypoperfusion, may also bedriving this remodelling. In this study, Iam examining the effects of this reducedblood supply on cortical organisation.The study involves magnetic resonanceimaging in a group of patients witharterial narrowing not causing stroke, orasymptomatic carotid stenosis. Perfusionscanning is performed to assess thearterial supply, MR spectroscopy to lookfor neuronal markers of chronic ischaemia,and functional MRI to create maps ofmotor and visual activation. These mapsare compared both with those obtainedfrom a healthy control group and theunaffected side in the patient population.If shifts to these activation maps aredemonstrated and correlated withperfusion imaging, these resultsmay provide evidence of how thebrain “prepares” itself for further injury,by shifting cortical function to other,well perfused regions of the functionalcortical network.The non-uniform topographic evolutionof the ischemic penumbra with time.Ma H, Zavala JA, Teoh H1, ChurilovL, Gunawan M, Ly J, Wright P, Phan T,Arakawa S, Davis S.M and Donnan G.AThe classical pattern of the ischemicpenumbra is defined by a central infarctcore surrounded by a uniform annulus ofsalvageable tissue. With time, numerousfactors may influence the rate and extentof penumbral salvage and, hence, alterthis pattern. We hypothesized that withco-registration of magnetic resonancediffusion and perfusion weighted images(MR DWI/PWI), we could identify aseries of differing penumbral patternswhich evolved with time. Patients wererecruited with MR studies performedwithin 48 hours of ischemic stroke onset.Mismatch pattern was demonstratedby co-registration of DWI/PWI imagesand categorized as classical (majority ofthe DWI within the PWI lesion) or nonclassicalpatterns. The proportion of thetwo patterns was assessed with referenceto time. Within 48 hours of stroke onsetboth classical and non-classical mismatchpatterns were identified. The formeroccurred earlier with larger mismatchvolumes. The reduction of classicalpattern with time reflected a non-uniformevolution of the ischemic penumbra.The recognition of these patternshighlights the need to individualizepatients when considering the potentialfor tissue salvage.Detection of small vessel knock signalsby Doppler ultrasound in a laboratorymodel simulating a penetrating branchocclusionChesda Udommongkol, Brian Chambers,Richard Manasseh, Ilija Sutalo, Ben Aldham,Emma Chung and Geoffrey DonnanSmall vessel knock (SVK) was recentlyreported as a new transcranial Doppler(TCD) sign of penetrating artery occlusion.However, knock-like signals may becaused by wall motion of normal basal109

Stroke— Stroke Imaging and Ultrasoundcerebral arteries. Preliminary in vitrostudies in Leicester suggest that SVK canbe recorded using TCD insonation of aside-branch lumen independent from thevessel wall.Aim: To demonstrate SVK in a laboratorymodel simulating a cerebral branchocclusion.Materials and Method: Our systemconsisted of a flow rig, peristaltic pump,reservoir tank and water tank. The flowcircuit was filled with blood mimickingfluid (BMF) that contained physical andacoustic properties similar to humanblood. Flow and viscosity were controlledto conform to normal human physiology.A scaled-up synthetic bifurcation wasmade from silicone tubing with internaldiameter 9.53 mm for the main arteryand 2.38 mm for the side branch. Itwas submerged in the water tank andconnected to the flow rig. The smallerbranch was insonated before and afterocclusion using a Compumedics DopplerBox with 2 MHz probe and Sonix RPUltrasonix with 7.5 MHz probe.Results: BMF recirculation in theblocked branch was detected by videorecording after dye injection and colourDoppler. Spectral Doppler demonstratedcorresponding continuous, bidirectional,low-frequency Doppler signals. HoweverSVK was not observed.Conclusion: In our in-vitro model, SVKwas not detected by TCD and duplexultrasound. Factors that might explainthese negative results include angle ofvessel bifurcation, flow pressure andtube material.Staff and StudentsGeoffrey Donnan MBBS, MD, FRACP,FRCP (Edin)Associate Professor Brian Chambers MBBS,MD, FRACPAmy Brodtmann MBBS FRACP PhDYoshinari Nagakane MDRajinder Dhamija MBBS, MDCaspar Brekenfeld MDAlvaro Cervera PhD, Dip of Advanced Studies,Doctorate in Biopathology in Medicine, Grad Dipin Tropical Medicine and Hygiene, MDJohn Ly MBBS, FRACPHenry Ma MBBS, FRACPRamesh Sahathevan MD (UKM), MRCP(ire),M.Med(UKM)Daniel Liu AMS StudentDr Chesda Udommongkol MDDr Anne Abbott Honorary Research FellowConferences andPresentationsTranslational Research in Neuroscience,University of Lund Hospital, Lund,Sweden, May 31, 2009.What’s New in Stroke: “Penumbralselection for trials of therapy – readyor not?” World Congress of Neurology,Bangkok, October 24, 2009.Collaborative LinksNationalDr Richard MannasehCSIRO, AustraliaSmall vessel knockInternationalProf Marc HommelUniversity of Grenoble, FranceProf Greg AlbersStanford Stroke Centre,Stanford University Medical Centre, USAImaging parameters for theischemic penumbraDr Emma ChungLeicester, UKSmall vessel knockDr Paul SymeScotland, UKSmall vessel knockEditorial PositionsProf Geoffrey DonnanStrokeJournal of NeuroimagingJournal of Internal MedicineLancet NeurologyInternational Journal of Stroke(Editor in Chief)Journal of the CardioMetabolic SyndromeAnnals of Indian Academy of NeurologyA/Prof Brian ChambersInternational Journal of Stroke110

PublicationsBrodtmann A, Puce A, Darby D, DonnanG. Regional fMRI brain activation doescorrelate with global brain volume. BrainRes 2009 Mar;1259:17-25.Brodtmann A, Puce A, Darby D, DonnanG. Serial Functional Imaging PoststrokeReveals Visual Cortex Reorganization.Neurorehabil Neural Repair 2009Feb;23(2):150-9.Davis SM, Donnan GA. 4.5 HoursThe New Time Window for TissuePlasminogen Activator in Stroke.Stroke 2009 Jun;40(6):2266-7.De Silva DA, Fink JN, Christensen S,Ebinger M, Bladin C, Levi CR, Parsons M,Butcher K, Barber PA, Donnan GA, DavisSM, Echoplanar Imaging Thrombolytic E.Assessing Reperfusion and Recanalizationas Markers of Clinical OutcomesAfter Intravenous Thrombolysis in theEchoplanar Imaging ThrombolyticEvaluation Trial (EPITHET). Stroke 2009Aug;40(8):2872-4.Donnan GA, Davis SM. AngiotensinReceptor Blockers and Stroke Therapy ItIs All About the Blood Pressure. Stroke2009 Sep;40(9):3163-.Ebinger M, Christensen S, De Silva DA,Parsons MW, Levi CR, Butcher KS, BladinCF, Barber PA, Donnan GA, Davis SM,Investigators E. Expediting MRI-BasedProof-of-Concept Stroke Trials Using anEarlier Imaging End Point. Stroke 2009Apr;40(4):1353-8.Ebinger M, Iwanaga T, Prosser JF, De SilvaDA, Christensen S, Collins M, ParsonsMW, Levi CR, Bladin CF, Barber PA,Donnan GA, Davis SM, Investigators E.Clinical-Diffusion Mismatch and BenefitFrom Thrombolysis 3 to 6 Hours AfterAcute Stroke. Stroke 2009 Jul;40(7):2572-4.Iwanaga T, Arakawa S, Siritho S, Fitt G,Dewey HM, Chambers BR, Donnan GA.Paracentral Strip Infarcts of the MiddleCerebral Artery: Borderzone Ischaemia orCortical Artery Occlusion? CerebrovascDis 2009;27(3):215-22.Levi C, Lindley R, Smith B, Bladin C,Parsons M, Read S, Cadilhac D, DonnanG, Davis S, Hankey G, Crimmins D,Gerraty R, Blacker D, Schultz D, GrimleyR, Lalor E, Natl Stroke Fdn Stroke Soc A.The implementation of intravenous tissueplasminogen activator in acute ischaemicstroke – a scientific position statementfrom the National Stroke Foundation andthe Stroke Society of Australasia. InternMed J 2009 May;39(5):317-24.Ma H, Zavala JA, Teoh H, ChurilovL, Gunawan M, Ly J, Wright P, PhanT, Arakawa S, Davis SM, Donnan GA.Fragmentation of the Classical MagneticResonance Mismatch “Penumbral”Pattern With Time. Stroke 2009Dec;40(12):3752-7.Ma H, Zavala JA, Teoh H, ChurilovL, Gunawan M, Ly J, Wright P, PhanT, Arakawa S, Davis SM, Donnan GA.Penumbral mismatch is underestimatedusing standard volumetric methods andthis is exacerbated with time. J NeurolNeurosurg Psychiatry 2009 Sep;80(9):991-6.Phan TG, Donnan GA, Srikanth V, ChenJ, Reutens DC. Heterogeneity in InfarctPatterns and Clinical Outcomes FollowingInternal Carotid Artery Occlusion. ArchNeurol 2009 Dec;66(12):1523-8.Phan TG, Fong AC, Donnan GA, SrikanthV, Reutens DC. Digital ProbabilisticAtlas of the Border Region between theMiddle and Posterior Cerebral Arteries.Cerebrovasc Dis 2009;27(6):529-36.Davis SM, Donnan GA. MR Mismatch andThrombolysis Appealing but ValidationRequired. Stroke. [Editorial Material]. 2009Aug;40(8):2910-.Donnan GA. Stroke diagnosis andimaging: a symbiotic partnership. IntJ Stroke. [Editorial Material]. 2010Apr;5(2):61-.Donnan GA, Baron JC, Ma H, Davis SM.Penumbral selection of patients for trialsof acute stroke therapy. Lancet Neurol.[Review]. 2009 Mar;8(3):261-9.Ebinger M, De Silva DA, Christensen S,Parsons MW, Markus R, Donnan GA, etal. Imaging the penumbra - strategiesto detect tissue at risk after ischemicstroke. J Clin Neurosci. [Review]. 2009Feb;16(2):178-87.111

Stroke— Neurorehabilitation and RecoveryOverviewOur aim within the division ofNeurorehabilitation and Recovery is toconduct and disseminate internationallycompetitiveresearch focussed oninvestigation of the scientific foundationsof neurological rehabilitation post-stroke.We address four major research themeswithin the division: (i) Mechanisms ofRecovery and Optimal Targeting ofRehabilitation; (ii) Restorative Approachesto Rehabilitation; (iii) Investigation ofthe Nature of Sensorimotor Impairmentand its Impact on Function; and (iv)Identification of Patients ‘at risk’ ofDepression through Novel Brain Imagingand Biomarkers.2009 has been an exciting and productiveyear for the Division, involving finaldata collection and ongoing analysisfor two major studies: IN_Touch(Imaging Neuroplasticity of Touch);and our randomised controlled trialSENSe (Study of the Effectiveness ofNeurorehabilitation on Sensation). Weare now embarking on two new projects:CoNNECT (Connecting New Networksfor Everyday Contract through Touch)and START (Stroke ImAging, pReventionand Treatment), leading to a number ofnew collaborations. We have continuedto contribute to international projects andhad prominent international scientists visitthe Division.An important feature of our program isthe collaborative links with researchersand centers locally and internationally.Collaborations span a number ofcountries and research groups includingUSA, Germany, France, UK, Sweden,Canada and New Zealand. In 2009we developed a new collaborativeresearch project focused on predictionof depression using imaging andcerebrovascular biomarkers over timein stroke. As part of this project weestablished collaborations with: DrOlivier Salvado, director of the e-Healthdivision of CSIRO, who has expertisein neuroimaging; Dr Thomas Linden,a neurologist and psychiatrist fromSweden with expertise in depressionpost-stroke; Prof Marc Hommel and DrAssia Jaillard, neurologists from Francewho have expertise in neuroimaging,depression and cogntive function poststrokeand Professor Sheila Crewther,a neuropsychologist who leads theNeuroscience program and CognitiveNeuroscience laboratory at La TrobeUniversity and has expertise in biomarkersand brain imaging.We are also expanding our collaborationsin rehabilitation science. Internationalcollaborations have commenced withthe Cognitive Rehabilitation ResearchGroup (CCRG), in St Louis, USA. TheCCRG is a major interdisciplinary researchgroup focused on improving methods ofcognitive rehabilitation for people withstrokes and involves a stroke registryof more than 10,000 patients. ProfessorCarey met with Professor Carolyn Baum,Director of the CCRG, in Houston, USA toplan collaborative projects on the impactof stroke and depression on activityparticipation.In the sensorimotor imaging field wehave continued to develop three newcollaborations, with new projects andgrant applications being submitted.We formed the Australasian StrokeRehabilitation and Imaging Scienceconsortium, ASTRIS. Members includeneurologists, rehabilitation scientists,movement scientists, and imaging expertsfrom Melbourne, Victoria; Newcastle,New South Wales; and New Zealand.Our collaboration with Dr Assia Jaillardand Prof Marc Hommel, neurologistsfrom Grenoble, France, has continued todevelop. Finally we have strengthenedour links with the Neuroimaging Divisionof FNI, with Dr Maria Gavrilescu andProf Gary Egan, as well as with Prof AlanConnelly and his team.We are involved, as experts, in twospecial international projects. TheNational Institute of Health (NIH)Toolbox: Assessment of Neurological andBehavioural Function project is part of theNIH Blueprint for Neuroscience Researchsupported by a US $21million grant.In 2009 we contributed to validationstudies in the area of somatosensoryassessment with healthy controls acrossthe lifespan and with stroke survivors.We are also involved in the JamesS Macdonnell collaborative project:Cognitive Neuroscience Principles forRehabilitation. The project involvesa select international group that willdevelop a text on translation of coreprinciples from cognitive neurosciencesto clinical practice. Prof Carey is the teamleader for the rehabilitation panel of theTouch, Space and Body Awareness group.Congratulations to Ettie Ben-Shabatand Melinda Randall who were awardedtheir PhDs in 2009. Ettie’s thesis is titled‘Central processing of proprioception:Functional MRI and psychophysicalstudies in healthy and stroke participants.’Melinda’s is titled ‘Modification andinvestigation of the construct validityof the Melbourne Assessment ofUnilateral Upper Limb Function’. Bothhave made major contributions to theirfield. Professor Leeanne Carey wasmade a Member of the Academy ofResearch of the American OccupationalTherapy Foundation for ‘exemplary anddistinguished contributions towards thescience of occupational Therapy. ProfCarey is the first Occupational Therapistin Australia to be inducted into theAcademy. Professor Carey was alsosuccessful in being awarded an AustralianResearch Council (ARC) Future Fellowshipin 2009.Major Research ProjectsIN_Touch: Imaging Neuroplasticity ofTouch. Brain adaptation associatedwith spontaneous and training-inducedrecovery of touch sensation post-stroke,using serial fMRI.Leeanne Carey (FNI), David Abbott (FNI),Rudiger Seitz (Germany), Aina Puce (USA)and Geoffrey Donnan (FNI)Brain networks may reorganise tooptimize stroke recovery. However,little is known about brain mechanismsunderlying spontaneous and traininginducedrecovery post-stroke, particularlyfollowing loss of body sensations. Weconducted serial functional brain imagingstudies in stroke survivors with loss oftouch sensation, such as ability to feeleveryday objects, and in healthy, agedmatched controls.In healthy controls reproducible activationwas identified in primary and secondarysensory regions of the brain associatedwith performance of a simple sensorydiscrimination task. Different patterns ofbrain activation were identified in strokesurvivors compared to controls. We foundnovel evidence of neural correlates ofpost-stroke touch impairment involvinga distributed network of ipsilesional SIand SII, contralesional thalamus, andattention regions, particularly followingsubcortical lesions. Findings suggest a112

modulatory role for ipsilesional SI andfrontal attention regions in processing oftouch. Rehabilitation directed at attentivediscrimination of sensory stimuli, ratherthan bombardment, is indicated.SENSe: Study of the Effectivenessof Neurorehabilitation on SensationLeanne Carey (FNI), Thomas Matyas(La Trobe University) and RichardMacdonell (Austin Health)After stroke loss of body sensationsis common, with negative impacton exploration of the immediateenvironment, hand function and returnto daily activities. Yet, there are currentlyfew approaches to sensory retrainingdescribed and limited controlledevidence of the effectiveness ofthese approaches. We developed aninnovative approach to rehabilitate loss ofsensation and hand function commonlyexperienced after stroke and tested itseffectiveness tested in a randomisedcontrol trial of 50 stroke survivors withvarying severity of sensory impairment.Between-group comparisons revealeda significantly greater improvementin sensory capacity following sensorydiscrimination training. Improvementswere clinically significant and maintainedat 6-weeks and 6-months. Patients withvarying side of lesion, severity of sensoryimpairment, age and interval post-strokewere able to benefit from training. Thisapproach to rehabilitation aims toimprove lost abilities rather than focuson compensation alone. The programis clinically-oriented and achievedtransfer of training effects to novelstimuli. Therapists can use this trainingapproach when working with people whoexperience sensory loss after stroke.Other Research ProjectsCoNNECT: Connecting New Networksfor Everyday Contact through Touch –pilot studyNew therapies have been developedto help the brain recover after stroke.Yet currently we do not have effectivemeans of identifying individuals whohave potential to benefit from thesetherapies. We will develop a model ofsensory recovery to predict the potentialfor recovery, and the most optimalrehabilitation for a stroke survivor.Brain imaging techniques will be usedto identify regions in the brain that arefunctionally and structurally connected.The ultimate goal of this study is to helptherapists choose optimal rehabilitationintervention for an individual, based onevidence of residual brain networks.PrePARE – Prediction and Preventionto Achieve Optimal RecoveryEndpoints after stroke: A substudyof STroke imAging pRevention andTreatment (START)Leeanne Carey (FNI), Sheila Crewther(LaTrobe), Thomas Linden (Sweden),Olivier Salvado (CSIRO) and MarcHommel (France)PrePARE is a new study that is designedto identify stroke surivors ‘at risk’ ofdeveloping post-stroke depression. Itis a prospective longitudinal study ofimaging, biochemical and geneticbiomarkers of post-stroke depression.The study is part of the START projectwith the overall aim to: i) determine thepredictive value of single and combinedbiochemical, genetic and imagingmarkers of post-stroke depression andrecovery; ii) determine if thrombolysis, anacute treatment known to be effectivein reducing disability, is associatedwith a reduced prevalence and severityof depression; iii) identify biologicalmechanisms underlying the developmentof post-stroke depression and recurrentstroke over 12 months. Our approachinvolves a multi-pronged and longitudinalapproach to investigating predictivemarkers of depression post-stroke.Depression will be assessed at acute,sub-acute and chronic phases. In thesubgroup of participants involved in thePrePARE study we will use advancedimaging techniques to investigate:i) the relationship between novel imagingmarkers (resting state activity, corticalthickness and fibre tract integrity) andfunctional outcome and depression andii) the association between post-strokedepression and functional outcomesincluding cognition, sensorimotorfunction, and participation in dailyactivities and life roles.Central processing of proprioception:fMRI and psychophysical studiesof healthy and stroke affectedparticipantsEttie Ben-Shabat, Leeanne Carey,Thomas Matyas and Amy BrodtmannProprioception is an essential componentof motor control, particularly for preciseupper limb movements. Despite itsimportance, knowledge of centralprocessing of proprioception and itspsychophysical function is lacking. Thethree main aims were to investigate:the psychophysical function ofproprioception; brain activation duringproprioceptive perception; and thechanges of both in stroke subjects withproprioceptive impairment. Functionalmagnetic resonance imaging scanswere performed during a cognitiveproprioceptive perception task. Theright supramarginal gyrus was activatedregardless of the stimulated wrist,providing a neuroanatomical explanationfor the observed higher accuracy ofleft compared to right proprioceptiveperception. Additional proprioceptivebrain activation was found in thehemisphere contralateral to the stimulatedhand, and included primary sensorymotor areas and the premotor cortex.Three case studies of stroke subjects withproprioceptive impairment and commonlesion site in posterolateral thalamus, wereconducted. Patterns of brain activationlacked the extent of right supramarginalactivation observed in healthy participants,and psychophysical function showeda clear deviation compared to healthyparticipants. Insights gained can assist inchoosing and interpreting psychophysicalproprioceptive tests. Combined withbrain activation studies, knowledgegained can improve diagnosis of subjectswith proprioceptive deficits, and aid infuture development of proprioceptivetreatments, together with the assessmentof their effectiveness. This study wasconducted as a PhD research thesis byEttie Ben-Shabat.113

Stroke— Neurorehabilitation & RecoveryThe contribution of somatosensoryimpairment to pinch grip abilityafter strokeJannette Blennerhassett (Austin Health),Leeanne Carey (FNI) and Thomas Matyas(NSRI, LaTrobe)Altered ability to grip and lift objectsis a major problem following stroke.Lack of controlled evidence aboutthe characteristics and underlyingmechanisms limit effective management.The aims of this study were to: a)characterise impairment on a controlledPinch Grip Lift and Hold (PGLH) taskwhen guided by somatosensoryinput, and b) investigate the relativecontribution of somatosensory and motordeficits to PGLH impairment post-stroke.Relative to matched healthy controls(n=45), PGLH impairment post-stroke(n=45) was characterised by prolongedgrip-lift phases, use of larger and morefluctuating force, and reduced grip-safetymargin adaptation to guard againstslip. Impairment was summarised bytwo orthogonal variables: Grip-LiftLatency and Grip Force Dysregulation.Poorer surface friction discriminationwas significantly associated with longergrip-lift latency and larger grip forcedysregulation during PGLH. Weightdiscrimination and visuomotor trackingwere not associated with PGLH deficit.Most stroke participants had abnormallyslowed dexterity and limited use of theirhands in routine daily tasks. The findingshighlight that somatosensory processingof friction between the skin and objectshould be integrated into trainingability to grip and lift objects followingstroke. This study was conducted asa PhD research thesis by Dr JannetteBlennerhassett.Additional Research ProjectsIN_Touch - late stages of recoveryLeeanne CareyNIH Toolbox: Assessment ofNeurological and Behavioural Function:SomatosensationLeeanne Carey, Winnie Dunn, (USA),Jamie Griffith (USA)Test-retest reliablity and normativestandards for the functional Test ofTactual Object Recognition, MultijointLimb position Sense Test etc.Leeanne CareyCognitive Neuroscience Principles forRehabilitation.Leeanne Carey, James S. McdonnellFoundationRecovery of touch sensationafter stroke: Attention, functionalconnectivity, and neuroplasticityLeeanne Carey, Sheila Crewther (La TrobeUniversity), Louise Bannister (La TrobeUniversity)A functional MRI study of upper limbtherapy in community dwelling strokesurvivorsIsobel Hubbard, Leeanne Carey,Mark ParsonsOccupation-based outcomes associatedwith sensory retraining post strokeMary Mastos (NSRI, LaTrobe), LeeanneCarey (NSRI, LaTrobe)An evaluation of occupationaltherapy and modified constraintinduced movement therapy followingBotulinum toxin-A injection in theupper limb in children with spastichemiplegic cerebral palsy.Brian Hoare (LaTrobe), Christine Imms(LaTrobe), Leeanne CareyModification and investigation of theconstruct validity of the MelbourneAssessment of Unilateral Upper LimbFunctionMelinda Randall (LaTrobe, Children’sHospital), Christine Imms (LaTrobe),Leeanne CareyFactors impacting on neural plasticityand recovery after stroke.Kate Noonan (LaTrobe), Sheila Crewther(LaTrobe), Leeanne CareyIs the Hand Function Survey reliableand responsive to change during strokerehabilitation?Rebecca Avery (LaTrobe). JannetteBlennerhassett (Austin Health, NSRI),Leeanne CareyRelationship between impairment,performance and participationmeasures post-strokeJacinta Spritzer, Tamara Tse, LeeanneCarey, Carolyn BaumFunctional and Structural ConnectivityChanges in the Brain in StrokeSurvivors with SomatosensoryImpairment: Implications forRehabilitationDavid McArdle, Leeanne Carey,Glynda KinsellaMechanisms by which inflammationmediates depression related celldeath, and the neuromodular effects ofimmune regulating nutritional factorsMichaela Pascoe, David Crewther, SheilaCrewther, Thomas Linden, Leeanne CareyImpact of depression post-stroke onactivity participation and quality of life.Tamara Tse, Leeanne Carey, JacintaDouglas, Primrose LentinDevelopment and implementationof a standardized sensory screeningtool for use with sub-acute patientspost stroke: Implementation of bestevidence into a clinical practice setting.Yvonne Mak, Leeanne Carey, Kate Richard114

Staff and StudentsLeeanne Carey BAppSc(OT), PhDMary Mastos BOTMatt Harvey BISTamara Tse BOTBonnie Alexander BSc, PGDipPsychDavid CareyHonorary AssociatesRudiger Seitz Honorary ProfessorThomas Matyas BA (Hons), PhDSheila Crewther BScHons, MSc,DipEd, BOptom, MAPrelimNeuropsych,PhD(Caltech), MAPSJacinta Douglas Honorary ProfessorAina Puce Honorary ProfessorMaria Gavrilescue Collaborator, FNIJannette Blennerhassett Honorary ResearchFellowCarolyn Baum Honorary ProfessorAssia Jaillard Honorary FellowStudentsEttie Ben-Shabat PhD CandidateMelinda Randall PhD CandidateBrian Hoare PhD CandidateIsobel Hubbard PhD CandidateMichaela Pascoe PhD CandidateKate Noonan Clinical DoctorateLouise Bannister Clinical DoctorateDavid McArdle Clinical DoctorateJohanne Walker Masters CandidateMary Mastos Masters CandidateJacinta Spitzer Honours CandidateYvonne Mak Honours CandidateConferences andPresentationsLeeanne CareyActivation Differs Markedly followingCortical or Subcortical Lesions CausingSomatosensory Impairment Post-Stroke.Organization for Human Brain Mapping15th Annual Meeting. June 18-23.San Francisco, CA.Sensory rehabilitation following stroke:Neuroscience foundations, new evidenceand application to clinical practice. Oneday workshop. OT Australia. AustralianAssociation of Occupational TherapistsVictoria. Melbourne Australia.Neuroscience Makes Sense forOccupational Therapy. Sadie PhilcoxLecture. University of Queensland.Media interactionsConnections: Occupational Therapy.Article on being awarded the Academyof Research Award from the AmericanOccupational Therapy FoundationABC radio interview: New treatmentsfor Stroke. Carol Duncan, Newcastle(12th May)ABC Health, The Pulse. Newtreatments for Stroke - Health andWellbeing. (16th April). – print (http://www.abc.net.au/health/thepulse/stories/2009/04/16/2544377.htm)Article New treatments for Stroke Health& Wellbeing in Australian PhysicianNational and InternationalCollaborative LinksProf Rudiger SeitzGermanyIN_Touch: Imaging Neuroplasticityof TouchProf Aina PuceUSAIN_Touch: Imaging Neuroplasticityof TouchProf Thomas MatyasLaTrobe University, AustraliaSENSe: Study of Effectiveness ofNeurorehabilitation on SensationProf Sheila CrewtherLaTrobe University, AustraliaPrePARE – Prediction and Preventionto Achieve Optimal Recovery Endpointsafter strokeProf Thomas LindenGothenberg, SwedenPrePARE – Prediction and Preventionto Achieve Optimal Recovery Endpointsafter strokeDr Olivier SalvadoCSIRO, AustraliaPrePARE – Prediction and Prevention toAchieve Optimal Recovery Endpointsafter strokeProf Marc HommelGrenobel, FrancePrePARE – Prediction and Preventionto Achieve Optimal Recovery Endpointsafter strokeProf Carolyn BaumUSAThe impact of sensory loss on activityparticipation post-strokeDr Assia JaillardGrenobel, FranceImaging motor recovery after strokeLaTrobe University Representativesfrom OT, Physiotherapy, Psychology,Communication Disorders and HumanBioscience, AustraliaPost-stroke rehabilitationAustralasian Stroke Rehabilitation andImaging Science consortium (ASTRIS)Members include neurologists,rehabilitation scientists, movementscientists, and imaging experts fromMelbourne, Victoria; Newcastle,New South Wales; and New ZealandProf Winnie DunnUSANIH Toolbox ProjectSusan Fitzpatrick and Maurizo CorbettaUSAJames S. Mcdonnell FoundationCognitive Neuroscience Principles forRehabilitation project115

Stroke— Neurorehabilitation & RecoveryEditorial PositionsPublicationsProf Leeanne CareyJournal of Neurorehabilitation and NeuralRepairOccupational Therapy InternationalAustralian Occupational Therapy JournalBrain Impairment (special issue)Visiting ScientistsDr Thomas Linden, SwedenDr Assia Jaillard and Professor MarcHommel, Grenobel, FranceAwardsLeeanne CareyMember of the Academy of Researchof the American Occupational TherapyFoundation.Australian Research Council,Future FellowshipEttie Ben-ShabatResearch Thesis Merit Citation,LaTrobe UniversityJohanne WalkerWindermere Foundation SpecialGrant -– ruralJacinta SpitzerJanet Sloan Stroke RehabilitationResearch AwardYvonne MakNational Stroke FoundationHonours AwardCarey, L.M. & Baum, C. (in press)Occupational Therapy. In Azari, N.P. (ed).Encyclopaedia of Sciences and Religions.Springer Verlag. Heidelberg, Germany.Hoare, B., Wallen, M., Imms, C.,Villanueva, E., Rawicki, H. B., Carey, L. (inpress) Botulinum toxin A as an adjunctto treatment in the management ofthe upper limb in children with spastichemiplegic cerebral palsy (CochraneReview): Update. In: The Cochrane Library.Chichester, UK: John Wiley & Sons, Ltd.Hoare, B., Imms, C., Rawicki, H. B., Carey,L. (in press) Modified constraint-inducedmovement therapy or conventionaloccupational therapy following injectionof Botulinum toxin-A to improve bimanualperformance in children with hemiplegiccerebral palsy: A randomised controlledtrial methods paper. BMC Neurology.Hubbard, I.J., Parsons, M.W., Neilson,C. & Carey, L.M. (2009). Task-specifictraining: Evidence for and translation toclinical practice. Occupational TherapyInternational. Vol 16 (3-4): 175-189.Pascoe, M., Crewther, D., Carey, L.,Crewther, S. (in press) Inflammation andDepression: Why Post Stroke DepressionMay be the Norm and not the Exception.International Journal of Stroke.Puce, A. & Carey, L. (in press).Somatosensory function. In Craighead,WE, Nemeroff, CB (Eds) The ConciseCorsini Encyclopedia of Psychology andBehavioral Science. 4th ed. New York.John Wiley & Sons, Inc.Spitzer, J., Tse, T., Baum, C., Carey, L.M. (inpress). Mild cognitive impairment impactson activity participation post-stroke in anAustralian cohort. Occupational TherapyJournal of Research: Occupational,Participation and Health.Other publications:Carey, L.M. & Linden, T. (2009)Assessment of Depression FollowingStroke. LaTrobe University (DVD.)Carey, L.M. & Linden, T. (2009) Impactof Sensory Loss Post-stroke. LaTrobeUniversity (DVD.)116

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Systems Neurophysiology— Brain Function in Health and DiseaseResearch OverviewProf Robin McAllen and Prof RichardMacdonell head the SystemsNeurophysiology Division at FNI. Ourresearch area is brain function in healthand disease. One particular focus ison how the brain controls basic bodilyfunctions such as blood pressure, bodytemperature, body fluids and breathing.A second focus is on the ways thatdisease processes change the excitabilityof neurons. A third focus is on the heart,and its complex interactions with thenervous system in health and disease.Major Research ProjectsChanges in motor cortex excitabilityin patients with epilepsy and responseto anticonvulsant treatmentBadawy RA, Macdonell RAL, Jackson GDand Berkovic SFThe electrical changes in the brain whichgive rise to epilepsy remain poorlyunderstood. Using transcranial magneticstimulation (TMS) we have shown thatthe increased excitability of the motorcortex occurs in both sides of the brainin generalised epilepsies whereas thishyperexcitability is confined to the sidecontaining the seizure focus in partialepilepsies. Using this technique we havealso shown that environmental factorssuch as sleep deprivation and timing ofthe menstrual cycle which are known toincrease seizure risk do so by increasingcortical excitability. After making thisfinding we were interested in exploringwhether seizure control using antiepilepticdrugs (AED) could be predicted using thistechnique. The TMS findings in patientswho successfully responded to drugsreturned to normal within a few weeksbut in patients where AEDs failed themotor cortex remained hyperexcitablesuggesting that this is a tool whichcould be used to test for likely ndividualresponsiveness to an AED or in thedevelopment of new drugs for epilepsy.We also explored the possible clinicalrole of TMS in assessing patientsafter a 1st seizure with respect to therisk of further seizures (epilepsy) anddemonstrated there is a correlationbetween increased cortical excitabilityand future risk of seizures. In addition wewere able to show that cortical excitabilityin epilepsy is a dynamic phenomenonwhich increases in the 24 hours beforea seizure, is reduced for 24 hours posta seizure before returning to a baselinehyperexcitable level compared withnormal controls . This wind up phase priorto a seizure might also potentially betherapeutic target if techniques with wideapplicability can be developed to identifyit prior to seizure onset. Manuscriptsdescribing this work have been publishedin international journals with the highestimpact factors in the field of neurology(Brain and Annals of Neurology) and theresults of the work are now being beentranslated into clinical care. Dr Badawyhas been awarded a Chancellor’s Prizefrom the University of Melbourne andcommendation in the 2010 Premiersaward for Health and Medical Researchfor her contribution.Brain regulation of body temperatureTanaka, M, McAllen RM and McKinley MJThe brain regulates body temperature,keeping it within a narrow range. Theconsequences when this fails can belife-threatening: elderly people, whosetemperature regulation may be impaired,frequently die of heat stroke duringheat waves.The master temperature controller regionin the brain is called the preoptic area.Neurons in this region adjust the balancebetween heat generation and heat lossfrom the body, using nerve connectionsto regulate skin blood flow, sweatingand heat generation by specialized fatdeposits (brown fat). If temperature fallstoo low, it also initiates shivering. Thesemechanisms set body temperature,but not always to an identical level.Our temperature is adjusted to about1 degree lower at night, for example,and upwards by one or two degrees ifwe are sick and run a fever. We also knowthat these nerve cells in the preopticregion take account not only of braintemperature but also of skin temperature.This combined information allows themto anticipate thermal challenges ratherthan simply reacting to them.What has become apparent from ourresearch over the past few years is that thebrain pathways controlling the differentdefences against losing heat in the cold –shutting down skin blood flow, generatingheat by brown fat or shivering – seem towork in parallel, quite independently ofeach other. This may seem surprising ifwe expect the brain to be engineeredas we would design it, but it seems lessstrange if we consider that each of thesemechanisms would have been adoptedat a different time in evolutionary history,and each would have been conservedbecause it independently helped theanimal to survive and reproduce. All thatis necessary for this arrangement to workis that each cold-defence response isengaged at an appropriate threshold skinor core temperature. Their brain pathwaysdo not need to communicate directly witheach other.118

This figure shows our current workingmodel, sketched on an outline of the rat’sbrain. We infer that each pathway hasits own temperature-sensitive neurons inthe preoptic area: those controlling skinblood vessels are exquisitely sensitive tobrain temperature, while those that driveshivering are much more responsive tocold skin; those regulating brown fat areintermediate. The motor output pathwaysfor all these actions have a synaptic relaystage in a brainstem nucleus call themedullary raphé, but despite this, theydo not appear to communicate witheach other. We are currently identifyingthe neural pathways that regulate skinblood flow.Sympathetic nerves in heart failureRohit Ramchandra, Sally Hood andClive MayHeart failure continues to be a majorhealth problem as evidenced by a risein hospital admissions for heart failure,the number of deaths attributed toheart failure, and the increasing costsassociated with care. It is established thatthe increase in activity of the nerves thatstimulate the heart to beat harder andfaster (cardiac sympathetic nerves) areactivated very strongly in heart failure.This overactivity exacerbates the diseaseprocess and can trigger sudden death.The activity of these nerves is controlledby the brain and our aim is to understandwhy they are overactive in heart failure.We have developed an animal model ofheart failure (the sheep) in which we haveshown by direct electrical recordings thatthere is a large increase in activity of thecardiac sympathetic nerves. We haveshown that in the healthy state activity inthese nerves is decreased by an increasein blood volume. It is thought that thisresults from increased pressures insidethe heart stimulating neural signals tothe brain, which reduces nerve activityto the heart.Understanding this is important becauseone of the consequences of heart failureis an increase in blood volume. We haverecently discovered that in heart failurethis system is dramatically desensitisedand increases in blood volume do notreduce cardiac sympathetic nerve activity.This indicates that in heart failure theinhibitory mechanisms that should inhibitcardiac sympathetic nerve activity inresponse to the increased blood volumeare absent, thus allowing other stimuli tomaintain activity at a high level.This graph shows how the activity in thesympathetic nerves to the heart (cardiacsympathetic nerves) respond to bloodvolume expansion. In normal sheep(closed symbols) volume receptors in theatria of the heart stimulate a reflex thatinhibits the activity of cardiac sympatheticnerves. This reflex stops working in sheepthat have heart failure (open symbols).119

Systems Neurophysiology— Brain Function In Health & DiseaseTreatment of Heart AttacksColleen Thomas, Bevyn Jarrott andClive May, in collaboration with ProfOwen Woodman (Discipline of CellBiology and Anatomy, RMIT University)and Dr Spencer Williams (School ofChemistry and Bio21 Molecular Scienceand Biotechnology Institute)Following an ischaemic event such as aheart attack or stroke, blood flow andthus oxygen transport to the downstreamtissue stops and the tissue dies. In tissuearound the ischaemic area blood flow andoxygen delivery are reduced, which leadsto further cell death and expansion ofthe damage from the original infarct. Theprimary intervention in the emergencydepartment is to remove the blockageand reperfuse the ischaemic tissue toprotect this area, which otherwise willeventually die. However reperfusionitself causes cell death and there areno treatments available to prevent thisreperfusion injury.We are developing a novel drugthat we have shown can reduce thereperfusion damage to the heart aftera heart attack by up to 30%, but theseexperiments were only over 3 hoursof reperfusion. In recent experimentswe have importantly confirmed thatthis beneficial cardioprotective effectis sustained over a period of 7 days.Furthermore we demonstrated thatthe impairment in the function of thecoronary arteries that occurs after a heartattack was also prevented by this drugtreatment. This drug could not be givenintravenously because it could only bedissolved in organic solvent, so we havedeveloped a water soluble pro-drugthat can be given intravenously and isequally cardioprotective. Further studiesare being conducted with the aim ofdeveloping this pro-drug as a clinicaltreatment for reperfusion injury.PublicationsBadawy RA, Macdonell RA, JacksonGD, Berkovic SF. Why do seizures ingeneralized epilepsy often occur in themorning? Neurology 2009;73(3):218-22.Badawy R, Macdonell R, Jackson G,Berkovic S. The peri-ictal state: corticalexcitability changes within 24 h of aseizure. Brain 2009;132(Pt 4):1013-21.Badawy RA, Harvey AS, MacdonellRA. Cortical hyperexcitability andepileptogenesis: Understanding themechanisms of epilepsy - part 2. J ClinNeurosci 2009;16(4):485-500.Badawy RA, Harvey AS, MacdonellRA. Cortical hyperexcitability andepileptogenesis: understanding themechanisms of epilepsy - part 1. J ClinNeurosci 2009;16(3):355-65.Chen D, Hazelwood L, Walker LL, OldfieldBJ, McKinley MJ, Allen AM. Changesin angiotensin type 1 receptor bindingand angiotensin-induced pressorresponses in the rostral ventrolateralmedulla of angiotensinogen knockoutmice. Am J Physiol Regul Integr CompPhysiol;298(2):R411-8.Chen N, Bezzina R, Hinch E,Lewandowski PA, Cameron-SmithD, Mathai ML, et al. Green tea, blacktea, and epigallocatechin modify bodycomposition, improve glucose tolerance,and differentially alter metabolic geneexpression in rats fed a high-fat diet. NutrRes 2009;29(11):784-93.Denton DA, McKinley MJ, Farrell M, EganGF. The role of primordial emotions inthe evolutionary origin of consciousness.Conscious Cogn 2009;18(2):500-14.Evans RG, May CN. Tissue hypoxia as atherapeutic target in acute kidney injury.Clin Exp Pharmacol Physiol 2009.Frithiof R, Ramchandra R, Hood S,May C, Rundgren M. Hypothalamicparaventricular nucleus mediates sodiuminducedchanges in cardiovascular andrenal function in conscious sheep. AmJ Physiol Regul Integr Comp Physiol2009;297(1):R185-93.Gall SL, Dewey HM, Sturm JW, MacdonellRA, Thrift AG. Handicap 5 years afterstroke in the North East MelbourneStroke Incidence Study. Cerebrovasc Dis2009;27(2):123-30.May CN, Evans RG. Frontiers inresearch series: Neural, hormonal andrenal interactions in long-term bloodpressure control II. Introduction. Clin ExpPharmacol Physiol;37(2):272-3.May CN, Frithiof R, Hood SG, McAllenRM, McKinley MJ, Ramchandra R. Specificcontrol of sympathetic nerve activity tothe mammalian heart and kidney. ExpPhysiol;95(1):34-40.McAllen RM, Tanaka M, Ootsuka Y,McKinley MJ. Multiple thermoregulatoryeffectors with independent centralcontrols. Eur J Appl Physiol;109(1):27-33.McAllen RM. Short of air? Cool it! JPhysiol 2009;587(Pt 21):5009-10.McKinley MJ, Weissenborn F, MathaiML. Drinking-induced thermoregulatorypanting in rehydrated sheep: influencesof oropharyngeal/esophageal signals,core temperature, and thirst satiety. AmJ Physiol Regul Integr Comp Physiol2009;296(6):R1881-8.Montano N, Furlan R, Guzzetti S, McAllenRM, Julien C. Analysis of sympatheticneural discharge in rats and humans.Philos Transact A Math Phys Eng Sci2009;367(1892):1265-82.Mookerjee I, Hewitson TD, Halls ML,Summers RJ, Mathai ML, Bathgate RA,et al. Relaxin inhibits renal myofibroblastdifferentiation via RXFP1, the nitricoxide pathway, and Smad2. Faseb J2009;23(4):1219-29.Parmar A, Langenberg C, Wan L, May CN,Bellomo R, Bagshaw SM. Epidemiologyof septic acute kidney injury. Curr DrugTargets 2009;10(12):1169-78.Prowle JR, Ishikawa K, May CN, Bellomo R.Renal blood flow during acute renal failurein man. Blood Purif 2009;28(3):216-25.Ramchandra R, Wan L, Hood SG, FrithiofR, Bellomo R, May CN. Septic shockinduces distinct changes in sympatheticnerve activity to the heart and kidneyin conscious sheep. Am J Physiol RegulIntegr Comp Physiol 2009;297(5):R1247-53.Ramchandra R, Hood SG, Frithiof R, MayCN. Discharge properties of cardiacand renal sympathetic nerves andtheir impaired responses to changesin blood volume in heart failure. AmJ Physiol Regul Integr Comp Physiol2009;297(3):R665-74.120

Ramchandra R, Hood SG, Denton DA,Woods RL, McKinley MJ, McAllen RM,et al. Basis for the preferential activationof cardiac sympathetic nerve activity inheart failure. Proc Natl Acad Sci U S A2009;106(3):924-8.Salo LM, Nalivaiko E, AndersonCR, McAllen RM. Control of cardiacrate, contractility, and atrioventricularconduction by medullary raphe neuronsin anesthetized rats. Am J Physiol HeartCirc Physiol 2009;296(2):H318-24.Tanaka M, McKinley MJ, McAllen RM.Roles of two preoptic cell groups in tonicand febrile control of rat tail sympatheticfibers. Am J Physiol Regul Integr CompPhysiol 2009;296(4):R1248-57.Thrift AG, Dewey HM, Sturm JW,Srikanth VK, Gilligan AK, Gall SL, etal. Incidence of stroke subtypes in theNorth East Melbourne Stroke IncidenceStudy (NEMESIS): differences betweenmen and women. Neuroepidemiology2009;32(1):11-8.Uschakov A, McGinty D, Szymusiak R,McKinley MJ. Functional correlates ofactivity in neurons projecting from thelamina terminalis to the ventrolateralperiaqueductal gray. Eur J Neurosci2009;30(12):2347-55.Wan L, Langenberg C, Bellomo R, MayCN. Angiotensin II in experimentalhyperdynamic sepsis. Crit Care2009;13(6):R190.Wang S, Thomas CJ, DustingGJ, Woodman OL, May CN.3’,4’-Dihydroxyflavonol improves postischaemiccoronary endothelial functionfollowing 7days reperfusion in sheep.Eur J Pharmacol 2009;624(1-3):31-7.121

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Florey Neuroscience InstitutesPARKVILLE CAMPUSLevel 2, Alan Gilbert Building161 Barry StreetCarlton South, VictoriaAustralia 3053Phone: +61 3 8344 1888Fax: +61 3 9347 0446AUSTIN CAMPUSNeuroscience BuildingAustin Health300 Waterdale RoadHeidelberg West, VictoriaAustralia 3081Phone: +61 3 9496 4137Fax: +61 3 9496 4071www.fni.edu.auFlorey Neuroscience Institutes is a publiccompany limited by guarantee under theCorporations Act 2001.ABN 92 124 762 027